Novel tyrosine kinase inhibitors

ABSTRACT

The present invention provides compounds of formula I  
                 
and pharmaceutically acceptable salts thereof. The formula I compounds inhibit tyrosine kinase enzymes thereby making them useful as anti-cancer agents.

RELATED APPLICATIONS

This application is a divisional of U.S. Ser. No. 10/751,798 filed Jan.5, 2004, which claims priority benefit under Title 35 § 119(e) of U.S.Provisional Application No. 60/437,926, filed Jan. 3, 2003, the contentsof which are incorporated by reference.

FIELD OF INVENTION

The present invention relates generally to the field of tyrosine kinaseenzyme inhibition using novel small molecules.

BACKGROUND OF THE INVENTION

Tyrosine Kinases are a class of enzymes, which catalyze the transfer ofthe terminal phosphate of adenosine triphosphate to the phenolichydroxyl group of a tyrosine residue present in the target protein.Tyrosine kinases play a critical role in signal transduction for severalcellular functions including cell proliferation, carcinogenesis,apoptosis, and cell differentiation (Plowman, G. D.; Ullrich, A.;Shawver, L. K.: Receptor Tyrosine Kinases As Targets For DrugIntervention. DN&P (1994) 7: 334-339). Therefore inhibitors of theseenzymes would be useful for the treatment or prevention of proliferativediseases which are dependent on these enzymes. Strong epidemiologicevidence suggests that the overexpression or activation of receptorprotein tyrosine kinases leading to constitutive mitogenic signaling isan important factor in a growing number of human malignancies. Tyrosinekinases that have been implicated in these processes include Abl, CDK's,EGF, EMT, FGF, FAK, Flk-1/KDR, HER-2, IGF-1R, IR, LCK, MEK, MET, PDGF,Src, and VEGF (Traxler, P. M. Protein Tyrosine Kinase Inhibitors inCancer Treatment. Exp. Opin. Ther. Patents (1997) 7: 571-588;incorporated herein by reference). Hence, there is an ongoing need toinvestigate novel compounds that can be used to regulate or inhibittyrosine kinase enzymes.

SUMMARY OF THE INVENTION

The present invention relates to compounds which inhibit tyrosine kinaseenzymes, compositions which contain tyrosine kinase inhibiting compoundsand methods of using inhibitors of tyrosine kinase enzymes to treatdiseases which are characterized by an overexpression or upregulation oftyrosine kinase activity such as cancer, diabetes, restenosis,arteriosclerosis, psoriasis, angiogenic diseases and immunologicdisorders (Powis, G.; Workman, P. Signaling targets For The Developmentof Cancer Drugs. Anti-Cancer Drug Design (1994), 9: 263-277; Merenmies,J.; Parada, L. F.; Henkemeyer, M. Receptor Tyrosine Kinase Signaling inVascular Development. Cell Growth Differ (1997) 8: 3-10; Shawver, L. K.;Lipsosn, K. E.; Fong, T. A. T.; McMahon, G.; Plowman, G. D.; Strawn, L.M. Receptor Tyrosine Kinases As Targets For Inhibition of Angiogenesis.Drug Discovery Today (1997) 2: 50-63; all herein incorporated byreference).

In addition to being used as single agents, it is contemplated thattyrosine kinase inhibitors can enhance the activity of cytotoxic orcytostatic treatments when used in combination with standard therapiesknown in the art.

The present invention is directed to compounds having Formula I

its enantiomers, diastereomers, pharmaceutically acceptable salts,hydrates, prodrugs and solvates thereof;wherein

A, B, D, and E are each, independently, C or N provided that if A, B, D,and E are each C, then one of R² and R³, R³ and R⁴, or R⁴ and R⁵ istaken together to form a heterocyclic ring having at least one nitrogenatom;

X is selected from the group consisting of N or C wherein each of said Nor C may be optionally substituted, independenty, with R⁷ and n is 0, 1,2, or 3;

Y is selected from the group consisting of O and S;

W is selected from the group consisting of N, C, O, and S, provided thatwhen W is O or S, R⁴¹ is absent;

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁴¹, and R⁴² are each independently selectedfrom the group consisting of H, C₁₋₆ alkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, halo, amino, aminoalkyl, alkoxy,thioalkoxy, nitro, aryl, heteroaryl, alkoxyalkyl, thioalkoxyalkyl,aminoalkyl, aralkyl, heteroarylalkyl, heterocycloalkylalkyl, —CN,—CO₂R⁸, —CONR⁹R¹⁰, —CO₂NR¹¹R¹², —NR¹³CONR¹⁴R¹⁵, —NR¹⁶SO₂R¹⁷,—SO₂NR¹⁸R¹⁹, —C(NR²⁰)NR²¹R²², —NH-Z, NH-Z-aryl, and NH-Z-heteroaryl, orany two of R² and R³, R³ and R⁴, or R⁴ and R⁵ can be taken together toform a heterocyclic ring having at least one nitrogen atom;

Z is selected from the group consisting of C₁-C₆ alkyl, cycloalkyl,alkenyl, cycloalkenyl, and alkynyl; Z optionally having one or morehydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR²³SO₂R²⁴ groups; Zoptionally incorporating one or more groups selected from the groupconsisting of —CO, —CNOH, —CNOR²⁶, —CNNR²⁷, —CNNCOR²⁸ and —CNNSO₂R²⁹;

R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹, R²²,R²³, R²⁴, and R²⁶ are independently selected from the group consistingof H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy,alkoxy, aryl, heteroaryl, heterocyclyl, heteroarylalkyl, and alkyl-R²⁵

wherein R²⁵ is alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino,alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, heteroaryl,heterocyloalkyl, sulfoxy, sulfonyl, —NR²⁷COOR²⁸, —NR²⁹C(O)R³⁰,—NR³¹SO₂R³², SO₂NR³¹R³², —C(O)NR³³R³⁴, and;

R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³ and R³⁴ are, independently, hydrogen,alkyl, or cycloalkyl.

The invention also provides a pharmaceutical composition comprising acompound of formula I, as defined above, and a pharmaceuticallyacceptable carrier.

The invention further provides a pharmaceutical composition comprising acompound of formula I, as defined above, in combination withpharmaceutically acceptable carrier and at least one other anti-canceragent optionally formulated as a fixed dose.

Additionally provided is a method of treating a condition associatedwith at least one tyrosine kinase enzyme comprising administering to amammalian species in need of such treatment an effective amount of acompound of formula I, as defined above. Furthermore, the inventionprovides a method of treating a condition associated with at least onetyrosine kinase enzyme comprising administering to a mammalian speciesat least one other anti-cancer agent in combination with a compound offormula I, as defined above.

The invention also provides for methods for treating proliferativediseases, such as cancer, comprising administering to a mammal in needof such treatment a therapeutically effective amount of a compoundhaving Formula I.

DESCRIPTION

The present invention provides for compounds of formula I, as definedabove, pharmaceutical compositions employing such compounds and methodsof using such compounds.

Listed below are definitions of various terms used to describe thecompounds of the instant invention. These definitions apply to the termsas they are used throughout the specification (unless they are otherwiselimited in specific instances) either individually or as part of alarger group.

The term “alkyl” herein alone or as part of another group refers to amonovalent alkane (hydrocarbon) derived radical containing from 1 to 12carbon atoms unless otherwise defined. An alkyl group is an optionallysubstituted straight, branched or cyclic saturated hydrocarbon group. Astraight chain alkyl group preferably contains from 1 to 6 carbon atoms.When substituted, alkyl groups may be substituted with up to foursubstituent groups, at any available point of attachment. When the alkylgroup is said to be substituted with an alkyl group, this is usedinterchangeably with “branched alkyl group”. Exemplary alkyl groupsinclude methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl,pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl,2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, and the like.Exemplary substituents may include but are not limited to one or more ofthe following groups: hydroxy, halo (such as F, Cl, Br, I), haloalkyl(such as CCl₃ or CF₃), alkoxy, alkylthio, cyano, carboxy (—COOH),alkylcarbonyl (—C(O)R), alkoxycarbonyl (—OCOR), amino, carbamoyl(—NHCOOR or —OCONHR), urea (—NHCONHR), thiol, (—SH), sulfoxy, sulfonyl,aryl, heteroaryl, and heterocycloalkyl. Alkyl groups as defined may alsocomprise one or more carbon to carbon double bonds or one or more carbonto carbon triple bonds. Alkyl groups may also be represented by theformula alkyl-R²⁵. In preferred embodiments, the alkyl group is amethyl, ethyl, propyl or butyl group and includes substituted methyl,ethyl, propyl or butyl groups.

The term “alkenyl” herein alone or as part of another group refers to ahydrocarbon radical straight, branched or cyclic containing from 2 to 12carbon atoms and at least one carbon to carbon double bond. An alkenylgroup may be optionally substituted in the same manner as described foran alkyl group.

The term “alkynyl” herein alone or as part of another group refers to ahydrocarbon radical straight, branched or cyclic containing from 2 to 12carbon atoms and at least one carbon to carbon triple bond. An alkynylgroup may be optionally substituted in the same manner as described foran alkyl group.

The term “alkoxy” as used alone or in combination herein refers to astraight or branched chain alkyl group covalently bonded to the parentmolecule through an oxygen atom linkage containing from one to tencarbon atoms and the terms “C₁₋₆ alkoxy” and “lower alkoxy” refer tosuch groups containing from one to six carbon atoms. Examples include,but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy,t-butoxy and the like. The term “optionally substituted” when used inconnection with an alkoxy substituent refers to the replacement of up totwo hydrogens, preferably on different carbon atoms with a radicalselected from the group of lower alkyl, phenyl, cyano, halo,trifluoromethyl, nitro, hydroxy, alkanoyl, amino, monoalkyl amino anddialkylamino. Alkoxy groups may be substituted in the same manner thatalkyl groups can be substituted as described above.

The term “sulfoxy” herein alone or as part of a group refers to —SO andmay be substituted with, for example, alkyl, aryl or heteroaryl groups.

The term “sulfonyl” herein alone or as part of a group refers to —SO₂and may be substituted with alkyl, aryl or heteroaryl groups.

The term “amino” herein alone or as part of another group refers to—NH₂. An “amino” may optionally be substituted with one or twosubstituents, which may be the same or different, such as alkyl, aryl,arylalkyl, alkenyl, alkynyl, heteroaryl, heteroarylalkyl,cycloheteroalkyl, cycloheteroalkylalkyl, cycloalkyl, cycloalkylalkyl,halo alkyl, hydroxyalkyl, alkoxyalkyl or thioalkyl. Preferredsubstituents include alkylamino and dialkylamino, such as methylamino,ethylamino, dimethylamino, and diethylamino. These substituents may befurther substituted with a carboxylic acid or any of the alkyl or arylsubstituents set out herein. In addition, the amino substituents may betaken together with the nitrogen atom to which they are attached to form1-pyrrolidinyl, 1-piperidinyl, 1-azepinyl, 4-morpholinyl,4-thiamorpholinyl, 4-sulfoxymorpholine, 4-sulfonylmorpholine,1-piperazinyl, 4-alkyl-1-piperazinyl, 4-arylalkyl-1-piperazinyl,4-diarylalkyl-1-piperazinyl, 1-homopiperazinyl,4-alkyl-1-homopiperazinyl, 4-arylalkyl-1-homopiperazinyl,4-diarylalkyl-1-homopiperazinyl; 1-pyrrolidinyl, 1-piperidinyl, or1-azepinyl, optionally substituted with alkyl, alkoxy, alkylthio, halo,trifluoromethyl or hydroxy.

The term “aryl” herein alone or as part of another group refers tomonocyclic or bicyclic aromatic rings, e.g. phenyl, substituted phenyland the like, as well as groups which are fused, e.g., napthyl,phenanthrenyl and the like. An aryl group thus contains at least onering having at least 6 atoms, with up to five such rings being present,containing up to 22 atoms therein, with alternating (resonating) doublebonds between adjacent carbon atoms or suitable heteroatoms. Aryl groupsmay optionally be substituted with one or more groups including, but notlimited to halogen, alkyl, alkenyl, alkynyl, alkoxy, hydroxy, carboxy,carbamoyl, alkyloxycarbonyl, alkylaminocarbonyl, nitro, trifluoromethyl,amino, cycloalkyl, cyano, alkyl S(O)_(m) (m=O, 1, 2), or thiol. Arylgroups may also be substituted with heterocycloalkyl and heterocycloarylgroups to form fused rings, such as dihydrobenzfuranyl, oxindolyl,indolyl, indolinyl, oxindolyl, benzoxazolidinonyl, benzoxazolinyl andbenzoxazolidinone.

The term “cycloalkyl” herein alone or as part of another group refers tofully saturated and partially unsaturated hydrocarbon rings of 3 to 9,preferably 3 to 7 carbon atoms. Further, a cycloalkyl may besubstituted. A substituted cycloalkyl refers to such rings having one,two, or three substituents, preferably one, selected from the groupconsisting of halo, alkyl, substituted alkyl, alkenyl, alkynyl, nitro,cyano, oxo (═O), hydroxy, alkoxy, thioalkyl, —CO₂H, —OC(═O)H, CO₂-alkyl,—OC(═O)alkyl, ═N—OH, ═N—O-alkyl, aryl, heteroaryl, heterocyclo, a fiveor six membered ketal (i.e. 1,3-dioxolane or 1,3-dioxane), —NR′R″,—C(═O)NR′R″, —OC(═O)NR′R″, —NR′CO₂′R″, —NR′C(═O)R″, —SO₂NR′R″, and—NR′SO₂R″, wherein each of R′ and R″ is independently selected fromhydrogen, alkyl, substituted alkyl, and cycloalkyl, or R′ and R″together form a heterocyclo or heteroaryl ring. Cycloalkyl groups mayalso be substituted with hetero atoms such as O, N, and S to formheterocycloalkyl groups. Preferred heterocycloalkyl groups includeoptionally substituted morpholine, homomorpholine (7 membered ring),thiomorpholine, piperazine, homopiperazine (7 membered ring), andpiperidine.

The term “heteroaryl” herein alone or as part of another group refers tosubstituted and unsubstituted aromatic 5 or 6 membered monocyclicgroups, 9 or 10 membered bicyclic groups, and 11 to 14 memberedtricyclic groups which have at least one heteroatom (O, S or N) in atleast one of the rings. Each ring of the heteroaryl group containing aheteroatom can contain one or two oxygen or sulfur atoms and/or from oneto four nitrogen atoms provided that the total number of heteroatoms ineach ring is four or less and each ring has at least one carbon atom.The fused rings completing the bicyclic and tricyclic groups may containonly carbon atoms and may be saturated, partially saturated, orunsaturated. The nitrogen and sulfur atoms may optionally be oxidizedand the nitrogen atoms may optionally be quatemized. Heteroaryl groupswhich are bicyclic or tricyclic must include at least one fully aromaticring but the other fused ring or rings may be aromatic or non-aromatic.The heteroaryl group may be attached at any available nitrogen or carbonatom of any ring. The heteroaryl ring system may contain zero, one, twoor three substituents selected from the group consisting of halo, alkyl,substituted alkyl, alkenyl, alkynyl, nitro, cyano, hydroxy, alkoxy,thioalkyl, —CO₂H, —OC(═O)H, —CO₂-alkyl, —OC(═O)alkyl, phenyl, benzyl,phenylethyl, phenyloxy, phenylthio, cycloalkyl, substituted cycloalkyl,heterocyclo, heteroaryl, —NR′R″, —C(═O)NR′R″, —OC(═O)NR′R″, —NR′CO₂′R″,—NR′C(═O)R″, —SO₂NR′R″, and —NR′SO₂R″, wherein each of R′ and R″ isindependently selected from hydrogen, alkyl, substituted alkyl, andcycloalkyl, or R′ and R″ together form a heterocyclo or heteroaryl ring.

Exemplary monocyclic heteroaryl groups include pyrrolyl, pyrrolidinyl,imidazolinyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl,thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thienyl, oxadiazolyl,pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, and the like.

Exemplary bicyclic heteroaryl groups include indolyl, indolinyl,oxindolyl, benzoxazolidinone, benzothiazolyl, benzodioxolyl,benzoxazolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl,isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl,chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl,indazolyl, pyrrolopyridyl, furopyridinyl, dihydroisoindolyl,tetrahydroquinolinyl and the like.

Exemplary tricyclic heteroaryl groups include carbazolyl, benzindolyl,phenanthrollinyl, acridinyl, phenanthridinyl, xanthenyl and the like.

The term “halogen” or “halo” herein alone or as part of another grouprefers to chlorine, bromine, fluorine or iodine selected on anindependent basis.

The term “hydroxy” herein alone or as part of another group refers to—OH.

The term “thioalkoxy” herein alone or as part of another group refers toan alkyl group as defined herein attached to the parent molecular groupthrough a sulfur atom. Examples of thioalkoxy include, but are notlimited to, thiomethoxy, thioethoxy, and the like.

Abbreviations: “Ph” represents phenyl; “Me” represents methyl; and “Et”represents ethyl.

An “anti-cancer agent” as used herein includes known anti-cancertreatments such as radiation therapy or with cytostatic or cytotoxicagents, such as for example, but not limited to, DNA interactive agents,such as cisplatin or doxorubicin; topoisomerase II inhibitors, such asetoposide; topoisomerase I inhibitors such as irinotecan or topotecan;tubulin interacting agents, such as paclitaxel, docetaxel or theepothilones; hormonal agents, such as tamoxifen; thymidilate synthaseinhibitors, such as 5-fluorouracil; anti-metabolites, such asmethotrexate; tyrosine kinase inhibitors such as Iressa and Tarceva;angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDKinhibitors; Her1/2 inhibitors and monoclonal antibodies directed againstgrowth factor receptors such as erbitux (EGF), herceptin (Her2), oravastin (VEGF).

When a functional group is termed “protected”, this means that the groupis in modified form to preclude undesired side reactions at theprotected site. Suitable protecting groups for the compounds of thepresent invention will be recognized from the present application takinginto account the level of skill in the art, and with reference tostandard textbooks, such as Greene, T. W. et al., Protective Groups inOrganic Synthesis, Wiley, N.Y. (1991).

When C₁₋₆ alkyl, alkenyl, alkynyl, cycloalkyl are substituted, they arepreferably substituted with one or more hydroxy, cyano, carbamoyl,hydroxy, alkoxy, thiol, alkenyl, thioalkoxy, amino, alkylamino, amido,sulfonyl, sulfoxy, sulfonamido, halo, heterocycloalkyl, aryl orheteroaryl.

When aryl or heteroaryl are substituted, they are preferably substitutedwith one or more alkyl, alkenyl, alkynyl, cyano, carbamoyl, hydroxy,alkoxy, thioalkoxy, amino, amido, sulfonamido, halo or with R′, R″wherein R′, R″ form a ring that is fused to the aryl group. When CH₂arylor heteroaryl are substituted, they are preferably substituted with oneor more alkyl, alkenyl, alkynyl, cyano, carbamoyl, hydroxy, alkoxy,thioalkoxy, amino, amido, sulfonamido, or halogen.

When NH-Z-aryl or NH-Z-heteroaryl groups are substituted, they arepreferably substituted with one or more alkyl, alkenyl, alkynyl,hydroxy, alkoxy, thioalkoxy, amino, halogen, nitro, nitrile,carboxylate, alkoxycarbonyl, carbamoyl, ester, amide, aryl, orheteroaryl groups.

The numbers in the subscript after the symbol “C” define the number ofcarbon atoms a particular group can contain. For example “C₁₋₆ alkyl”means a straight or branched saturated carbon chain having from one tosix carbon atoms; examples include methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl, t-butyl, n-pentyl, sec-pentyl, isopentyl,and n-hexyl. Depending on the context, “C₁₋₆ alkyl” can also refer toC₁₋₆ alkylene which bridges two groups; examples includepropane-1,3-diyl, butane-1,4-diyl, 2-methyl-butane-1,4-diyl, etc. “C₂₋₆alkenyl” means a straight or branched carbon chain having at least onecarbon-carbon double bond, and having from two to six carbon atoms;examples include ethenyl, propenyl, isopropenyl, butenyl, isobutenyl,pentenyl, and hexenyl. Depending on the context, “C₂₋₆ alkenyl” can alsorefer to C₂₋₆ alkenediyl which bridges two groups; examples includeethylene-1,2-diyl (vinylene), 2-methyl-2-butene-1,4-diyl,2-hexene-1,6-diyl, etc. “C₂₋₆ alkynyl” means a straight or branchedcarbon chain having at least one carbon-carbon triple bond, and from twoto six carbon atoms; examples include ethynyl, propynyl, butynyl, andhexynyl.

The term “alkyl-R²⁵” includes optionally substituted alkyl groups suchas methyl, ethyl, propyl, and butyl, attached to and R²⁵ group. R²⁵generally includes hydrogen, alkenyl, hydroxy, thiol, alkoxy,thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano,halo, sulfoxy, sulfonyl, —NHCOOH, —NHC(O)—, —NHSO₂—, —C(O)NH₂,heteroaryl or heterocycloalkyl groups such as morpholinyl or a grouphaving the formula:

wherein R₅₁ is H or alkyl.

The terms “imidazole” and “imidazoline” herein alone or as part ofanother group includes substituted imidazoles and substitutedimidazolines. Similarly, the term “tetrahydropyrimidine” includessubstituted tetrahydropyrimidines. Likewise, the terms “piperazine”,“piperidine” “morpholines”, “homopiperazines”, “homomorpholines” and“pyrrolidine” include substituted piperazines, substituted piperidines,substituted morpholines, substituted homomorpholines and substitutedpyrrolidines, respectively

Compounds of the present invention have the general formula I:

and include it enantiomers, diastereomers, pharmaceutically acceptablesalts, hydrates, prodrugs and solvates thereof;wherein

A, B, D, and E are each, independently, C or N;

X is selected from the group consisting of N or C wherein each of said Nor C may be optionally substituted, independenty, with R⁷ and n is 0, 1,2, or 3;

Y is selected from the group consisting of O and S;

W is selected from the group consisting of N, C, O, and S, provided thatwhen W is O or S, R⁴¹ is absent;

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁴¹, and R⁴² are each independently selectedfrom the group consisting of H, C₁₋₆ alkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, halo, amino, aminoalkyl, alkoxy,thioalkoxy, nitro, aryl, heteroaryl, alkoxyalkyl, thioalkoxyalkyl,aminoalkyl, aralkyl, heteroarylalkyl, heterocycloalkylalkyl, —CN,—CO₂R⁸, —CONR⁹R¹⁰, —CO₂NR¹¹R¹², —NR¹³CONR¹⁴R¹⁵, —NR¹⁶SO₂R¹⁷,—SO₂NR¹⁸R¹⁹, —C(NR²⁰)NR²¹R²², —NH-Z, —NH-Z-aryl, and NH-Z-heteroaryl, orany two of R² and R³, R³ and R⁴, or R⁴ and R⁵ can be taken together toform a heterocyclic ring having at least one nitrogen atom;

Z is selected from the group consisting of C₁-C₆ alkyl, cycloalkyl,alkenyl, cycloalkenyl, and alkynyl; Z having one or more hydroxy, thiol,alkoxy, thioalkoxy, amino, halo, NR²³SO₂R²⁴ groups; Z optionallyincorporating one or more groups selected from the group consisting of—CO, —CNOH, —CNOR²⁶, —CNNR²⁷, —CNNCOR²⁸ and —CNNSO₂R²⁹;

R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹, R²²,R²³, R²⁴, and R²⁶ are independently selected from the group consistingof H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy,alkoxy, aryl, heteroaryl, heterocyclyl, heteroarylalkyl, and alkyl-R²⁵

wherein R²⁵ is alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino,alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, heteroaryl,heterocyloalkyl, sulfoxy, sulfonyl, —NR²⁷COOR²⁸, —NR²⁹C(O)R³⁰,—NR³¹SO₂R³², SO₂NR³¹R³², —C(O)NR³³R³⁴, and;

R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³ and R³⁴ are, independently, hydrogen,alkyl, or cycloalkyl.

In some embodiments of the present invention, if A, B, D and E are eachC, then at least one of R² and R³, R³ and R⁴ or R⁴ and R⁵ is takentogether to form a heterocyclic ring. Preferred heterocyclic ringsinclude optionally substituted imidazole, pyrazole, pyridine,pyrimidine, pyrrolidine, morpholine, piperidine, and piperazine.

In preferred embodiments, R³ is an amino group such as NHCH₂CH₂OH,NMeCH₂CH₂OH, NEtCH₂CH₂OH, NHCH₂CH₂NH₂, NMeCH₂CH₂NH₂, NEtCH₂CH₂NH₂,NHCH₂CH₂NMe₂, NMeCH₂CH₂NMe₂, NEtCH₂CH₂NMe₂, NHCH₂CH₂NEt₂, NMeCH₂CH₂NEt₂,NEtCH₂CH₂NEt₂, NHCH₂CH₂N(CH₂CH₂)₂O, NMeCH₂CH₂N(CH₂CH₂)₂O, orNEtCH₂CH₂N(CH₂CH₂)₂O; a heteroaryl, a heterocyclo, or an alkoxy group,—OR, wherein R is H or alkyl-R²⁵ and R²⁵ is as defined above.

In some embodiments, R³ is an optionally substitued piperidine.Preferred substituents are selected from the group consisting ofhydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, 1,3dioxolane (—OCHR)₂, 1,3 dioxane (—OCHRCHRCHRO—) —NHC(O)R, —NHCO₂Rwherein R is alkyl or alkyl-R²⁵.

In some embodiments of the present invention, R³ is an optionallysubstituted morpholine, homomorpholine, thiomorpholine,sulfoxymorpholine, or sulfonylmorpholine. Preferred substituents includehydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy,alkyl-R²⁵, —NHC(O)R, —NHCO₂R, wherein R is hydrogen, alkyl or alkyl-R²⁵.

In some embodiments, R³ is a pyrrolidine. Preferred pyrrolidinesinclude, 3-hydroxyl pyrrolidine, 3-alkoxy pyrrolidine, and 3-alkylaminopyrrolidine.

In some embodiments, R³ is an optionally substitutedN-tetrahydropyrimidine or N-imidazoline wherein the substituents are,preferably, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cyanoalkyl,carboxyl, or carboxamide.

In some embodiments, R⁶ is is selected from the group consisting ofamino, NH-Z, NH-Z-aryl, and NH-Z-heteroaryl; wherein Z is defined asabove.

In preferred embodiments, R⁶ is is selected from the group consisting H,2-aminomethylpyridine, NHCH₂CH(OH)aryl, and NHCH(CH₂OH)CH₂aryl, whereinthe aryl group is optionally substituted. In preferred embodiments, thearyl group is substituted with at least one Br, Cl, F, or methoxy andmay have one of the following formulae:

wherein R⁴⁰ is hydrogen or alkyl, preferably methyl, and R⁷⁰ is hydrogenor halogen, such as Br, Cl or F.

Suitable examples of salts of the compounds according to the inventioninclude inorganic or organic acids. These include, but are not limitedto, hydrochloride, hydrobromide, sulfate, methanesulfonate, maleate,fumarate, phosphate and other pharmaceutically acceptable salts. Saltswhich are unsuitable for pharmaceutical uses but which can be employed,for example, for the isolation or purification of free compounds offormula I or their pharmaceutically acceptable salts, are also included.

All stereoisomers of the compounds of the instant invention arecontemplated, either in admixture or in pure or substantially pure form.The definition of the compounds according to the invention embraces allpossible stereoisomers and their mixtures. It very particularly embracesthe racemic forms and the isolated optical isomers having the specifiedactivity. The racemic forms can be resolved by physical methods, suchas, for example, fractional crystallization, separation orcrystallization of diastereomeric derivatives or separation by chiralcolumn chromatography. The individual optical isomers can be obtainedfrom the racemates by conventional methods, such as, for example, saltformation with an optically active acid followed by crystallization.

It should be understood that the present invention includes prodrugforms of the compounds of formula I. Various forms of prodrugs are wellknown in the art. For examples of such prodrug derivatives, see:

-   (a) Design of Prodrugs, edited by H. Bundgaard (Elsevier, 1985); and    Methods in Enzymology, Vol. 42, pp. 309-396, edited by K. Widder et    al., (Academic Press, 1985);-   (b) A Textbook of Drug Design and Development, edited by    Krosgaard-Larsen and H. Bundgaard, Chapter 5, “Design and    Application of Prodrugs,” by H. Bundgaard, pp. 113-191 (1991);-   (c) H. Bundgaard, Advanced Drug Deliver Reviews, 8, pp. 1-38 (1992);-   (d) H. Bundgaard et al., Journal of Pharmaceutical Sciences, 77, 285    (1988); and-   (e) N. Kayeka et al., Chem. Phar. Bull., 32, 692 (1984).

The invention also provides a pharmaceutical composition comprising acompound of formula I, as defined above, and a pharmaceuticallyacceptable carrier and at least one other anti-cancer agent formulatedas a fixed dose. Preferred anti-cancer agents are selected from thegroup consisting of: tamoxifen, toremifen, raloxifene, droloxifene,iodoxyfene, megestrol acetate, anastrozole, letrazole, borazole,exemestane, flutamide, nilutamide, bicalutamide, cyproterone acetate,goserelin acetate, luprolide, finasteride, herceptin, methotrexate,5-fluorouracil, cytosine arabinoside, doxorubicin, daunomycin,epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin,cisplatin, carboplatin, melphalan, chlorambucil, busulphan,cyclophosphamide, ifosfamide, nitrosoureas, thiotephan, vincristine,taxol, taxotere, etoposide, teniposide, amsacrine, Irinotecan,topotecan, an epothilone; a tyrosine kinase inhibitor such as Iressa orTarceva; an angiogenesis inhibitor; an EGF inhibitor; a VEGF inhibitor;a CDK inhibitor; a Her1/2 inhibitor and monoclonal antibodies directedagainst growth factor receptors such as erbitux (EGF) and herceptin(Her2).

The invention further provides a method of treating a condition viamodulation of at least one tyrosine kinase enzyme comprisingadministering to a mammalian species in need of such treatment aneffective amount of a compound of formula I, as defined above.

Additionally, the invention provides a method of treating a conditionvia modulation of at least one tyrosine kinase enzyme comprisingadministering to a mammalian species in need of such treatment aneffective amount of a compound of formula I, as defined above, incombination (simultaneously or sequentially) with at least one otheranti-cancer agent.

A preferred condition, treated by said methods of the instant invention,is cancer. Additionally, the tyrosine kinase enzyme may include (but isnot limited to): Abl, CDK's, EGF, EMT, FGF, FAK, Flk-1/KDR, HER-2,IGF-1R, IR, LCK, MEK, MET, PDGF, Src, and VEGF.

The invention also provides a method for treating cancer, comprisingadministering to a mammalian species in need of such treatment, atherapeutically effective amount of at least one of the pharmaceuticalcompositions defined above.

The invention further provides a method for treating proliferativediseases, comprising administering to a mammalian species in need ofsuch treatment a therapeutically effective amount of at least one of thepharmaceutical compositions defined above.

Certain compounds of formula I may generally be prepared according tothe following schemes and the knowledge of one skilled in the art.Solvates (e.g., hydrates) of the compounds of formula I are also withinthe scope of the present invention. Methods of salvation are generallyknown in the art. Accordingly, the compounds of the instant inventionmay be in the free or hydrate form, and may be obtained by methodsexemplified by the following schemes below.

Schemes I-IX illustrate the preparation of compounds claimed in thisinvention. The examples, which follow, illustrate the compounds that canbe synthesized by these schemes. The schemes are not limited by theexamples listed or by any substituents employed for illustrativepurposes.

Scheme I describes the preparation of the benzimidazoles. The startingdiamines 1 are readily available using literature methods or areobtained commercially. The diamine is then condensed with an aldehyde 2to provide the benzimidazole 3. Further modification of the functionalgroups appended to A-E, or R⁷, R⁴¹, or R⁴² are then possible.

Alternatively, the benzimidazole could be formed in a step-wise manner(see Scheme II and Scheme III). Scheme II describes the preparation ofthe benzimidazoles via the amide formation using the acid chloride of 5or any of the commonly used peptide coupling reagents such as DCC(dicyclohexylcarbodiimide), EDCI(1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), etc. Oncethe amide 6 is formed the nitro group is reduced using catalytichydrogenation, transfer hydrogenation or chemical reduction such asSnCl₂ or iron powder or other methods known in the art for reduction ofaryl nitro groups. Treatment of the aniline with acid then forms thebenzimidazole.

Alternatively, the diamine 1 could be used in the amide formingreaction. Such a diamine could be deprotoned with strong base such asLiHMDS or LDA followed by addition of the alkyl ester derivative of thesaid acid 5, preferably the methyl ester, to provide an intermediatewhich could be dehydrated preferably with POCl₃ to provide thebenzimidazole 3

Scheme III illustrates the condensation of the diamine 1 with analdehyde 2 to give the imine 7. Once the Schiff base 7 is formed thearylimine is induced to undergo oxidative cyclization using iodobenzenediacetate (IDB) as an oxidant to provide the benzimidazole.

For example, Scheme IV illustrates the use of4-iodo-2-methoxy-pyridine-3-carbaldehyde 8 to provide the functionalizedbenzimidazole 9. Hydrolysis of the methoxy group using protic acidconditions, TMSI (trimethylsilyl iodide), BBr₃, or other conditionsknown in the art for cleaving a methyl ether provides the halopyridone10. For some benzimidazoles where A,B,D or E is “N” hydroxylsis of themethoxy group results in cleavage of the benzimidazole ring to an aminoamide. This amide can be reclosed with POCl₃ or other dehydratingreagent to reclose the benzimidazole ring and provide halopyridone 9.Addition of heteroatom nucleophiles using amines, alcohols or thiolsthen provides the substituted pyridones 11. Other functionality could beincorporated into the aldehyde and the above example is included forillustrative purposes only.

Likewise the aryl ring of the benzimidazole prepared using Schemes I, IIor III can be modified. For example introduction of a cyano group for R³on the benzimidazole allows for the formation of heterocycles such asimidazole, imidazolines, oxazolines, thiazolines, amides, or amidines.Scheme V illustrates such transformations. Starting from thecyano-substituted benzimidazole 12 the heterocycle can be modified asillustrated in Scheme V to provide 13. Imidate formation preferablyusing ethanol and acid provides intermediate 14. Imidate 14 can betransformed using diamines to form imidazolines, amino alcohols to formoxazolines, amino acetals to form imidazoles, and amino thiols to formthiazolines 15. Alternatively the imidate can be hydrolyzed to the acidand coupled with amines using any of the standard amide formationreagents (DCC, EDCI, etc.) to form amides 16. Imidate 14 is also auseful intermediate for the preparation of amidines 17 by reacting withamines.

Scheme VI illustrates further transformation of benzimidazoles that beara halogen atom using palladium catalysis using conditions developed bySuzuki [Yang et al. Acta Chem. Scand. (1993) 221; Suzuki et al. Synth.Commun. (1981) 11: 513] or Buchwald/Hartwig [Buchwald et al. J. Am.Chem. Soc. (1994) 116: 7901; Hartwig et al. J. Am. Chem. Soc. (1994)116: 5969; Hartwig. Angew. Chem., Int. Ed. Engl. (1998) 37: 2046] andvariations of these methods. Preparation of a bromide substitutedbenzimidazole 18 is envisioned to provide a substrate for Suzukicoupling with aryl, vinyl, and heterocyclic boronic acids to providebenzimidazoles 19. Likewise, amines and heterocycles such as piperazineor morpholine derivatives 20 can be prepared from the same bromide usingamines under conditions described by Buchwald and Hartwig or variationsthereof.

Alternatively amine and heterocyclic derivatives such as 20 is preparedusing intermediate 6 described in Scheme II. When the R³ of 6 is ahalogen, preferably F, the halogen can be displaced with amines,alcohols, heterocyclic amines and other nitrogen containing heterocyclessuch as piperazine, piperidine, 4-amino piperidine, morpholine,imidazole, etc (Scheme VII). The terminal nitrogen of piperazine or4-amino piperidine is then alkylated using standard alkylationconditions or reacted with aldehydes in a reductive amination reactionto provide alkylated derivatives. Alternatively the terminal nitrogenatom of piperazine or 4-amino piperidine can be acylated orcarbamoylated using any number of conditions that are routine forsomeone skilled in the art of organic synthesis. Following the exampleillustrated in Scheme II compounds such as 20 could be prepared.

Alternatively amines, heterocycles, and alcohols can be introduced at R³using a nucleophilic aromatic substitution reaction started from anintermediate 21 were R³ is halogen, preferably Cl. The halogen can beactivated by either A and/or D being a N, or by being activated by ap-nitro group (NR¹ ₂; where R¹ is O). The halogen can be displaced withamines, alcohols, heterocyclic amines and other nitrogen containingheterocycles such as piperazine, piperidine, 4-amino piperidine,morpholine, imidazole, etc (Scheme VIII). The R¹2 and NR²2 can then bedeprotected or reduced (when R¹ or R² is O) to provide the diamine. Theterminal nitrogen of piperazine or 4-amino piperidine is then alkylatedusing standard alkylation conditions or reacted with aldehydes in areductive amination reaction to provide alkylated derivatives.Alternatively the terminal nitrogen atom of piperazine or 4-aminopiperidine is acylated or carbamoylated using any number of conditionsthat are routine for someone skilled in the art of organic synthesis.The resulting nitro or amino groups NR¹ ₂ and NR² ₂ are then deprotectedor reduced (when R¹ or R² is O) to provide the diamine 22 and processedas illustrated in Scheme III.

Scheme IX illustrates the synthesis of a specific heterocylicbenzimidazole for illustration. Starting from2,6-dichloro-3-nitro-pyridin-4-ylamine, 23 (R. J. Rousseau and R. K.Robins J. Heterocyclic Chem. 2 (1965), p. 196) a methyl group isintroduced using Pd catalysis and trimethylaluminium to provide pyridine24. The chloro group of 24 can be displaced by any number ofnucleophiles but amine nucleophiles are specfically envisioned such asmorpholine, piperazine, piperadine and pyrolidine and substitutedderivatives thereof. The resulting nitro aniline 25 can be reduced bycatalytic hydrogenation, transfer hydrogenation or chemical reductionsuch as SnCl₂ or iron powder or other methods known in the art forreduction of aryl nitro groups. The diamine can be 1′ can be isolated orcondensed crude with aldehyde 8 as illustrated in Scheme IV. FollowingScheme IV benzimidazole derivatives such as 11′ can be prepared.

Utility

The compounds according to the invention have pharmacologicalproperties; in particular, the compounds of formula I are tyrosinekinase enzyme inhibitors. The novel compounds of formula I are thususeful in the therapy of a variety of proliferative diseases (includingbut not limited to diseases associated with tyrosine kinase enzymes)such as cancer, autoimmune diseases, viral diseases, fungal diseases,neurodegenerative disorders and cardiovascular disease.

More specifically, the compounds of formula I are useful in thetreatment of a variety of cancers, including, but not limited to, thefollowing:

a) carcinoma, including that of the bladder, breast, colon, kidney,liver, lung, including small cell lung cancer, esophagus, gall bladder,ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, includingsquamous cell carcinoma;

b) hematopoietic tumors of lymphoid lineage, including leukemia, acutelymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma,T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy celllymphoma and Burkett's lymphoma;

c) hematopoietic tumors of myeloid lineage, including acute and chronicmyelogenous leukemias, myelodysplastic syndrome and promyelocyticleukemia;

d) tumors of mesenchymal origin, including fibrosarcoma andrhabdomyosarcoma;

e) tumors of the central and peripheral nervous system, includingastrocytoma, neuroblastoma, glioma and schwannomas; and

f) other tumors, including sarcoma, melanoma, seminoma, teratocarcinoma,osteosarcoma, xenoderoma pigmentosum, keratoctanthoma, thyroidfollicular cancer and Kaposi's sarcoma.

Due to the key role of tyrosine kinases in the regulation of cellularproliferation in general, inhibitors could act as reversible cytostaticagents which may be useful in the treatment of any disease process whichfeatures abnormal cellular proliferation, e.g., benign prostatichyperplasia, familial adenomatosis polyposis, neuro-fibromatosis,atherosclerosis, pulmonary fibrosis, arthritis, psoriasis,glomerulonephritis, restenosis following angioplasty or vascularsurgery, hypertrophic scar formation, inflammatory bowel disease,transplantation rejection, endotoxic shock, and fungal infections.

Compounds of formula I may induce apoptosis. The apoptotic response isaberrant in a variety of human diseases. Compounds of formula I, asmodulators of apoptosis, will be useful in the treatment of cancer(including but not limited to those types mentioned herein above), viralinfections (including but not limited to herpesvirus, poxvirus,Epstein-Barr virus, Sindbis virus and adenovirus), prevention of AIDSdevelopment in HIV-infected individuals, autoimmune diseases (includingbut not limited to systemic lupus, erythematosus, autoimmune mediatedglomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory boweldisease, and autoimmune diabetes mellitus), neurodegenerative disorders(including but not limited to Alzheimer's disease, AIDS-relateddementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitispigmentosa, spinal muscular atrophy and cerebellar degeneration),myelodysplastic syndromes, aplastic anemia, ischemic injury associatedwith myocardial infarctions, stroke and reperfusion injury, arrhythmia,atherosclerosis, toxin-induced or alcohol related liver diseases,hematological diseases (including but not limited to chronic anemia andaplastic anemia), degenerative diseases of the musculoskeletal system(including but not limited to osteoporosis and arthritis)aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple sclerosis,kidney diseases and cancer pain.

Compounds of formula I may modulate the level of cellular RNA and DNAsynthesis. These agents would therefore be useful in the treatment ofviral infections (including but not limited to HIV, human papillomavirus, herpesvirus, poxvirus, Epstein-Barr virus, Sindbis virus andadenovirus).

Compounds of formula I may also be useful in the chemoprevention ofcancer. Chemoprevention is defined as inhibiting the development ofinvasive cancer by either blocking the initiating mutagenic event or byblocking the progression of pre-malignant cells that have alreadysuffered an insult or inhibiting tumor relapse.

Compounds of formula I may also be useful in inhibiting tumorangiogenesis and metastasis.

The compounds of this invention may also be useful in combination(administered together or sequentially) with known anti-cancertreatments such as radiation therapy or with cytostatic or cytotoxicagents, such as for example, but not limited to, DNA interactive agents,such as cisplatin or doxorubicin; topoisomerase II inhibitors, such asetoposide; topoisomerase I inhibitors such as Irinotecan or topotecan;tubulin interacting agents, such as paclitaxel, docetaxel or theepothilones; hormonal agents, such as tamoxifen; thymidilate synthaseinhibitors, such as 5-fluorouracil; and anti-metabolites, such asmethotrexate; tyrosine kinase inhibitors such as Iressa and Tarceva;angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDKinhibitors; Her1/2 inhibitors and monoclonal antibodies directed againstgrowth factor receptors such as erbitux (EGF) and herceptin (Her2).

If formulated as a fixed dose, such combination products employ thecompounds of this invention within the dosage range described below andthe other pharmaceutically active agent or treatment within its approveddosage range. Compounds of formula I may also be administeredsequentially with known anticancer or cytotoxic agents when acombination formulation is inappropriate. The invention is not limitedin the sequence of administration; compounds of formula I may beadministered either prior to or after administration of the knownanticancer or cytotoxic agent(s).

Further subject matter of the invention also includes pharmaceuticalsfor use, as described above, including controlling cancer, inflammationand arthritis, which contain at least one compound of the formula I asdefined above or at least one of its pharmacologically acceptable acidaddition salts, and the use of a compound of the formula I as definedabove for the preparation of a pharmaceutical having activity againstproliferative diseases as described previously including against cancer,inflammation and/or arthritis.

The pharmacological properties of the compounds of this invention may beconfirmed by a number of pharmacological assays. The exemplifiedpharmacological assays which follow have been carried out with thecompounds according to the invention and their salts.

Biological Assays

A. CDK 2/Cyclin E Kinase Assay

Kinase reactions consisted of 5 ng of baculovirus expressedGST-CDK2/cyclin E complex, 0.5 μg GST-RB fusion protein (amino acids776-928 of retinoblastoma protein), 0.2 μCi ³³P γ-ATP, 25 μM ATP in 50μl kinase buffer (50 mM Hepes, pH 8.0, 10 mM MgCl₂, 1 mM EGTA, 2 mMDTT). Reactions were incubated for 45 minutes at 30° C. and stopped bythe addition of cold trichloroacetic acid (TCA) to a final concentrationof 15%. TCA precipitates were collected onto GF/C unifilter plates(Packard Instrument Co., Meriden, Conn.) using a Filtermate universalharvester (Packard Instrument Co., Meriden, Conn.) and the filters werequantitated using a TopCount 96-well liquid scintillation counter(Packard Instrument Co., Meriden, Conn.). Dose response curves weregenerated to determine the concentration required inhibiting 50% ofkinase activity (IC₅₀). Compounds were dissolved at 10 mM indimethylsulfoxide (DMSO) and evaluated at six concentrations, each intriplicate. The final concentration of DMSO in the assay equaled 2%.IC₅₀ values were derived by non-linear regression analysis and have acoefficient of variance (SD/mean, n=6)=14%.

B. EMT Kinase Assay

A filter-based kinase assay, measuring the phosphorylation of Gst-SLP76by Gst-Emtk, was employed to determine the compound inhibitory activityagainst Emt. The kinase reaction was performed in a 96-well plate atroom temperature for 15 min before being terminated by adding 100 μl of20% trichloroacetic acid (TCA) containing 0.1 M sodium pyrophosphate.The kinase reaction mixture (60 μl) contained 25 mM HEPES, pH 7.0, 0.1mg/ml BSA, 5 mM MgCl₂, 5 mM MnCl₂, 8 ng of enzyme (Gst-Emtk), 5 μg ofthe substrate protein (Gst-SLP76), 1 μM ATP, 0.4 μCi of [γ-P³³]ATP andthe tested compound (at various concentrations). After termination, theproteins were allowed to precipitate in the presence of TCA for 1 hr at4° C. The precipitated proteins were then harvested on a filter plate(UniFilter-96, GF/C, Packard Instrument) and washed to remove excess[γ-P³³]ATP. The radioactivity was determined using a TopCount NXT(Packard Instrument) after adding 35 μl of Microscint 20 (PackardInstrument).

C. FAK Tyrosine Kinase Assay

The Focal Adhesion kinase was assayed using the synthetic polymerpoly(Glu/Tyr) (Sigma Chemicals) as a phosphoacceptor substrate. Eachreaction mixture consisted of a total volume of 50 ul and contained 100ng of baculovirus-expressed enzyme, 2 μg of poly(Glu/Tyr), 1 μM of ATP,and 0.2 μCi of [γ-³³P]ATP. The mixtures also contained 40 mM Tris.HCl,pH 7.4, 1 mM MnCl₂, 0.5 mM DTT, and 0.1 mg/ml bovine serum albumin. Thereaction mixtures were incubated at 26° C. for 1 hour and kinaseactivity was determined by quantitation of the amount of radioactivephosphate transferred to the poly(Glu/Tyr) substrate. Incorporation wasmeasured by the addition of cold trichloroacetic acid (TCA)precipitation of the proteins which were collected onto GF/C unifilterplates (Packard Instrument Co., Meriden, Conn.) using a Filtermateuniversal harvester and the filters were quantitated using a TopCount96-well liquid scintillation counter (Packard Instrument Co., Meriden,Conn.). Compounds were dissolved in dimethyl sulfoxide to aconcentration of 10 mM and were evaluated at six concentrations, each intriplicate. The final concentration of DMSO added to the kinase assayswas 0.5%, which has been shown to have no effect on kinase activity.IC50 values were derived non-linear regression analysis and have acoefficient of variance (SD/mean, n=6)=16%.

D. HER-1/HER-2 Kinase Assay

Kinase reactions consisted of 10 ng of baculovirus expressed GST-HER1,100 ng of HER2, 100 ng/ml poly(Glu/Tyr) (Sigma), 0.2 μCi 33P γ-ATP, 1 μMATP in 50 μl kinase buffer (50 mM Tris, pH 7.5, 10 mM MnCl2, 0.5 mMDTT). Reactions were incubated for 1 h at 27 C and stopped by theaddition of cold trichloroacetic acid (TCA) to a final concentration15%. TCA precipitates were collected onto GF/C unifilter plates (PackardInstrument Co., Meriden, Conn.) using a Filtermate universal harvester(Packard Instrument Co., Meriden, Conn.) and the filters werequantitated using a TopCount 96-well liquid scintillation counter(Packard Instrument Co., Meriden, Conn.). Dose response curves weregenerated to determine the concentration required to inhibit 50% ofkinase activity (IC₅₀). Compounds were dissolved at 10 mM indimethylsulfoxide (DMSO) and evaluated at six concentrations, each intriplicate. The final concentration of DMSO in the assay equaled 1%.IC₅₀ values were derived by non-linear regression analysis and have acoefficient of variance (SD/mean, n=6)=16%.

E. IGF-Receptor Tyrosine Kinase Assay

The IGF-1 receptor tyrosine kinase was assayed using the syntheticpolymer poly(Glu/Tyr) (Sigma Chemicals) as a phosphoacceptor substrate.Each reaction mixture consisted of a total volume of 50 ul and contained125 ng of baculovirus expressed enzyme, 2.5 μg of poly(Glu/Tyr), 25 μMof ATP, and 0.1 μCi of [γ-³³P]ATP. The mixtures also contained 20 mMMOPS, pH 7.0, 5 mM MnCl₂, 0.5 mM DDT, and 0.1 mg/ml bovine serumalbumin. The reaction mixtures were incubated at 30° C. for 45 minutesand kinase activity was determined by quantitation of the amount ofradioactive phosphate transferred to the poly(Glu/Tyr) substrate.Incorporation was measured by the addition of cold trichloroacetic acid(TCA) precipitation of the proteins which were collected onto GF/Cunifilter plates (Packard Instrument Co., Meriden, Conn.) using aFiltermate universal harvester and the filters were quantitated using aTopCount 96-well liquid scintillation counter (Packard Instrument Co.,Meriden, Conn.). Compounds were dissolved in dimethyl sulfoxide to aconcentration of 10 mM and were evaluated at six concentrations, each intriplicate. The final concentration of DMSO added to the kinase assayswas 0.5%, which has been shown to have no effect on kinase activity.IC50 values were derived by non-linear regression analysis and have acoefficient of variance (SD/mean, n=6)=16%.

F. Insulin Receptor Tyrosine Kinase Assay

The Insulin Receptor Tryrosine kinase was assayed using the syntheticpolymer poly(Glu/Tyr) (Sigma Chemicals) as a phosphoacceptor substrate.Each reaction mixture consisted of a total volume of 50 ul and contained90 ng of baculovirus-expressed enzyme, 2.5 μg of poly(Glu/Tyr), 25 μM ofATP, and 0.1 μCi of [γ-³³P]ATP. The mixtures contained also 20 mMTris.HCl, pH 7.4, 5 mM MnCl₂, 0.5 mM DTT, and 0.1 mg/ml bovine serum.The reaction mixtures were incubated at 26° C. for 1 hour and kinaseactivity was determined by quantitation of the amount of radioactivephosphate transferred to the poly(Glu/Tyr) substrate. Incorporation wasmeasured by the addition of cold trichloroacetic acid (TCA)precipitation of the proteins which were collected onto GF/C unifilterplates (Packard Instrument Co., Meriden, Conn.) using a Filtermateuniversal harvester and the filters were quantitated using a TopCount96-well liquid scintillation counter (Packard Instrument Co., Meriden,Conn.). Compounds were dissolved in dimethyl sulfoxide to aconcentration of 10 mM and were evaluated at six concentrations, each intriplicate. The final concentration of DMSO added to the kinase assayswas 0.5%, which has been shown to have no effect on kinase activity.IC50 values were derived non-linear regression analysis and have acoefficient of variance (SD/mean, n=6)=16%.

G. LCK Kinase Assay

Kinase reactions consisted of 10 ng of baculovirus expressed 10 ngGST-Lck, 100 ng/ml poly(Glu/Tyr) (Sigma), 0.2 μCi 33P γ-ATP, 1 μM ATP in50 μl kinase buffer (50 mM Tris, pH 7.5, 10 mM MnCl2, 0.5 mM DTT).Reactions were incubated for 1 h at 27 C and stopped by the addition ofcold trichloroacetic acid (TCA) to a final concentration 15%. TCAprecipitates were collected onto GF/C unifilter plates (PackardInstrument Co., Meriden, Conn.) using a Filtermate universal harvester(Packard Instrument Co., Meriden, Conn.) and the filters werequantitated using a TopCount 96-well liquid scintillation counter(Packard Instrument Co., Meriden, Conn.). Dose response curves weregenerated to determine the concentration required to inhibit 50% ofkinase activity (IC₅₀). Compounds were dissolved at 10 mM indimethylsulfoxide (DMSO) and evaluated at six concentrations, each intriplicate. The final concentration of DMSO in the assay equaled 1%.IC₅₀ values were derived by non-linear regression analysis and have acoefficient of variance (SD/mean, n=6)=16%.

H. MET Kinase Assay

Kinase reactions consisted of 10 ng of baculovirus expressed GST-Met,2.5 ug poly(Glu/Tyr) (Sigma), 0.2 μCi 33P γ-ATP, 10 μM ATP in 50 μlkinase buffer (40 mM Tris, pH 7.5, 1 mM MnCl2, 0.50 mM DTT). Reactionswere incubated for 1 h at 27 C and stopped by the addition of coldtrichloroacetic acid (TCA) to a final concentration 3.5%. TCAprecipitates were collected onto GF/C unifilter plates (PackardInstrument Co., Meriden, Conn.) using a Filtermate universal harvester(Packard Instrument Co., Meriden, Conn.) and the filters werequantitated using a TopCount 96-well liquid scintillation counter(Packard Instrument Co., Meriden, Conn.). Dose response curves weregenerated to determine the concentration required to inhibit 50% ofkinase activity (IC₅₀). Compounds were dissolved at 10 mM indimethylsulfoxide (DMSO) and evaluated at seven concentrations, each intriplicate. The final concentration of DMSO in the assay equaled 1%.IC₅₀ values were derived by non-linear regression analysis and have acoefficient of variance (SD/mean, n=6)=16%.

I. PDGF Receptor Kinase Assay

Kinase reactions consisted of 70 ng of baculovirus expressed GST-PDGFbR,0.3 ug biotinylated poly(Glu/Tyr) (Sigma), in 50 μl kinase buffer (20 mMHepes, pH 7.5, 0.7 uM ATP, 10 mM MnCl2, 0.5 mM DTT, 0.15 mM NaCl, 0.1mg/ml BSA). Reactions were incubated for 30 minutes at room temperaturewith shaking and stopped by the addition of 10 ul of 0.2M EDTA, pH8.0.150 ul of HTRF detection buffer was added and incubated for 1 hour and30 minutes at room temperature. Counts were quantitated on DiscoveryHTRF Packard Instrument.

J. VEGFR-2 (KDR) Kinase Assay

Kinase reactions consisted of 7.5 ng of baculovirus expressed GST-KDR,1.5 ug poly(Glu/Tyr) (Sigma), 0.04 μCi 33P γ-ATP, 2.5 μM ATP in 50 μlkinase buffer (25 mM Tris, pH 7.5, 1.8 mM MnCl2, 0.0.625 mM DTT).Reactions were incubated for 1 h at 27 C and stopped by the addition ofcold trichloroacetic acid (TCA) to a final concentration 15%. TCAprecipitates were collected onto GF/C unifilter plates (PackardInstrument Co., Meriden, Conn.) using a Filtermate universal harvester(Packard Instrument Co., Meriden, Conn.) and the filters werequantitated using a TopCount 96-well liquid scintillation counter(Packard Instrument Co., Meriden, Conn.). Dose response curves weregenerated to determine the concentration required to inhibit 50% ofkinase activity (IC₅₀). Compounds were dissolved at 10 mM indimethylsulfoxide (DMSO) and evaluated at six concentrations, each intriplicate. The final concentration of DMSO in the assay equaled 1%.IC₅₀ values were derived by non-linear regression analysis and have acoefficient of variance (SD/mean, n=6)=16%.

K. Cytotoxicity Assay (HT-29-colon; Colo205, MCF-7-breast)

Tumor cell lines are maintained in McCoy's 5 A medium (GIBCO) and 10%heat inactivated fetal bovine serum (GIBCO). The in vitro cytotoxicityis assessed in tumor cells by a tetrazolium-based colorimetric assaywhich takes advantage of the metabolic conversion of MTS(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphenyl)-2H-tetrazolium,inner salt) (Promega) to a reduced form that absorbs light at 492 nm(1). Cells are seeded 24 hr prior to drug addition. Following a 72 hourincubation at 37° C. with serially diluted test compound, MTS (Riss, T.L, et al., Comparison of MTT, XTT, and a novel tetrazolium compound MTSfor in vitro proliferation and chemosensitivity assays.,” Mol. Biol.Cell 3 (Suppl.): 184a, 1992), in combination with the electron couplingagent phenazine methosulfate, is added to the cells. The incubation iscontinued for 3 hours, then the absorbency of the medium at 492 nm ismeasured with a spectrophotometer to obtain the number of survivingcells relative to control populations. The results are expressed asmedian cytotoxic concentrations (IC₅₀ values).

-   Biological Activity; the compounds of the present invention had    kinase activity of <25 uM against one or more of the following    kinases CDK, EMT, FAK, Her1, Her2, IGF, IR, LCK, MET, PDGF, VEGF.

General Procedure for the Preparation of2-Hydroxy-2-(Substituted-phenyl)-ethylamines

1-(3-Chloro-phenyl)-2-nitro-ethanol:To a solution of3-chloro-benzaldehyde (20 g, 0.142 mol) in nitromethane (100 mL) wereadded magnesium sulfate (37.6 g, 0.312 mol) and phosphazene baseP₁-t-bu-tris(tetramethylene) (4.43 g, 0.014 mol). The reaction mixturewas stirred at room temperature for 2 h. After concentration in vacuo,the residue was purified by flash chromatography (25% EtOAc/hexane) toyield the title compound (26.4 g, 100%) as a green-yellow oil. ¹H NMR(300 MHz, DMSO-d₆) δ 7.53 (1H, s), 7.35-7.42 (3H, m), 6.23 (1H, broads), 5.32-5.33 (1H, m), 4.90 (1H, dd, J=3.2, 12.4 Hz), 4.60 (1H, dd,J=1.2, 12.4 Hz).

[1-(3-Chloro-phenyl)-2-nitro-ethoxy]-triethyl-silane: To a solution of1-(3-chloro-phenyl)-2-nitro-ethanol (26 g, 0.14 mol) in DMF (50 mL) wereadded imidazole (28.6 g, 0.42 mol) and chlorotriethylsilane (25.3 g,0.17 mol). The reaction mixture was stirred at room temperature for 2 h.After quenching with water, the mixture was extracted with ethylacetate. The combined organic layers were washed with water and brine,dried over Na₂SO₄, and filtered. After removal of solvent, the crudeproduct was purified by flash chromatography (2% EtOAc/hexane) to yieldthe title compound (37 g, 91%) as a colorless oil. ¹H NMR (300 MHz,CDCl₃) δ 7.40 (1H, s), 7.27-7.32 (3H, m), 5.40 (1H, dd, J=3.2, 9.5 Hz),4.51 (1H, dd, J=9.5, 12.1 Hz), 4.36 (1H, dd, J=3.3, 12.1 Hz), 0.85 (9H,t, J=7.5 Hz), 0.54 (6H, q, J=7.5 Hz).

2-(3-Chloro-phenyl)-2-triethylsilanyloxy-ethylamine: Raney nickel (1 g)was washed with distilled water five times and methanol three times.[1-(3-Chloro-phenyl)-2-nitro-ethoxy]-triethyl-silane (10 g, 0.032 mol)and Raney nickel in methanol (100 mL) was hydrogenated (35 psi) at roomtemperature for 14 h. The reaction mixture was filtered through a pad ofcelite and rinsed with methanol. Concentration of the filtrate todryness gave the title compound (5.6 g, 62%) as a colorless oil whichwas used for the next step without purification. ¹H NMR (300 MHz, CDCl₃)δ 7.32 (1H, s), 7.18-7.26 (3H, m), 4.70 (1H, t, J=5.8 Hz), 2.86 (2H, m),0.89 (9H, t, J=7.9 Hz), 0.56 (6H, q, J=7.8 Hz). LRMS (M+H)⁺ m/z 286.

General Procedure for the Preparation of2-Hydroxy-2-(substituted-phenyl)-ethylamines

4-methoxy-3-bromophenyl chloroacetophenone: To a suspension of AlCl₃(13.4 g, 0.10 mol) in methylene chloride (40 mL) was added a solution of2-bromoanisole (12.5 mL, 0.10 mol) and chloroacetyl chloride (8 mL, 0.10mol) at 0° C. The solution was warmed to ambient temperature for twohours and poured onto ice and extracted with methylene chloride, washedwith saturated sodium bicarbonate solution, brine, and dried over MgSO₄.The solution was filtered, concentrated and crystalized from EtOH togive 15.37 g of white solid. LRMS [M−H]-260.8; IR (KBr) 1697, 1048, 1255cm⁻¹; ¹H NMR (300 MHz, CDCl₃) δ 8.18 (s, 1H), 7.94 (dd, J=8.67 Hz, 1H),6.96 (d, J=8.67 Hz, 1H), 4.62 (s, 2H), 3.98 (s, 3H); ¹³C NMR (CDCl₃,75.5 Hz) δ 188.8, 160.3, 134.1, 129.9, 128.2, 112.4, 111.3, 56.6, 45.3.

General Procedure for Chiral Reduction of Chloroketones and Ammonolysis

(S)-1-[4-methoxy-3-bromophenyl]-2-chloro ethanol: To a solution of(S)-Methyl-CBS-oxazaborolidine (1M in toluene, 0.745 mL, 0.745 mmol) andBH₃-THF (8 mL, 8 mmol) was added at the same time a solution of BH₃-THF(19 mL, 19 mmol) and a solution of the chloroketone (10.03 g, 37.98mmol) in 19 mL of THF. Both solutions were added dropwise over 30minutes. The solution was stirred for 1 hour and quenched with the slowaddition of methanol (50 mL). The solution was concentrated and theresidue chromatographed over a short silica gel column (1:1 hexane/ethylacetate) to give a quantitative yield (10.0 g) of chlorohydrin as aclear oil. IR (KBr) 1053, 1258, 3406 cm⁻¹; ¹H NMR (300 MHz, CDCl₃) δ7.59 (s, 1H), 7.30 (dd, J=2.16 Hz, 1H), 6.90 (d, J=8.46 Hz, 1H), 4.83(dd, J=3.57 Hz, 1H), 3.90 (s, 3H), 3.64 (ddd, J=3.6, 11.1, 8.7, 2H),2.04 (b s, 1H). ¹³C NMR (CDCl₃, 75.5 MHz) δ 155.9, 133.5, 131.1, 126.3,111.9, 73.1, 60.4, 56.3, 50.7.

(S) 2-Amino-1-[3-chloro-4-methoxyphenyl]ethanol Hydrochloride: To asolution of the chlorohydrin (10.0 g, 37.9 mmol) in 120 mL of methanolat −40° C. was added 100 grams of ammonia. The solution was sealed in apressure bottle and warmed to ambient temperature and stirred for 48hours. The solution was cooled and opened. The ammonia was allowed toevaporate and solution concentrated. The residue was crystalized fromethanol/ethyl acetate to give 3.83 g of white solid (35%). The materialwas reacted with Boc₂O in ethyl acetate and saturated sodium bicarbonateand analyzed by chiral HPLC using a chiralcel OJ column using 95%hexane/ethanol as elutant and determined to by 98% ee. Additional cropswere collected −2.96 g and 1.41 g for a total of 75% yield. LRMS[M+H]+246; IR (cm⁻¹, KBr) 1055, 1261, 3001, 2948, 3356; ¹H NMR (500 MHz,DMSO) δ 8.09 (b s, 2H), 7.58 (s, 1H), 7.36 (dd, J=2.05, 6.45 Hz, 1H),7.11 (d, J=8.5 Hz, 1H) 6.10 (s, 1H), 4.80 (m, 1H), 3.84 (s, 3H), 3.00(ddd, J=2.7, 12.6, 9.5 Hz, 2H); ¹³C NMR (DMSO, 75.5 MHz) δ 154.8, 135.4,130.4, 126.6, 112.4, 110.4, 67.9, 56.2, 45.4.

(S) 2-Amino-1-[3-chlorophenyl]ethanol Hydrochloride: was preparedaccording to the general procedure outlined above. LRMS [M+H]+172; IR(KBr, cm-1) 3048, 3351, 2952; ¹H NMR(300 MHz, MeOD) δ 7.48 (s, 1H), 7.35(m, 3H), 3.31 (ddd, J=1.5, 3.12, 9.15 Hz 2H).

(S)-2-Amino-1-[3-bromophenyl]ethanol Hydrochloride: was preparedaccording to the general procedure outlined above. LRMS [MH]+217.9; IR(KBr, cm-1) 3025, 3443, 2891; ¹H NMR (500 MHz, DMSO) δ 7.93 (b s, 2H),7.60 (s, 1H), 7.52 (d, 1H), 7.41 (s, 1H), 7.35 (d, J=7.7 Hz, 1H) 6.17(s, 1H), 4.82 (m, 1H), 3.08 (ddd, J=2.6, 12.7, 9.6 Hz, 2H); ¹³C NMR(DMSO, 75.5 MHz) δ 144.4, 30.5, 128.7, 125.0, 121.6, 68.3, 45.1.

(S)-2-Amino-1-[3-chloro-4-methylthiophenyl]ethanol Hydrochloride: wasprepared according to the general procedure outlined above. LRMS[M+H]+217.9; IR (KBr, cm-1) 3007, 3358; ¹H NMR (500 MHz, DMSO) δ 8.12 (bs, 2H), 7.46 (s, 1H), 7.37 (s, 1H), 7.35 (d, 1H) 6.19 (d, 1H), 4.83 (m,1H), 3.01 (ddd, J=3.2, 12.8, 9.3 Hz, 2H); ¹³C NMR (DMSO, 75.5 MHz) δ139.6, 136.5, 129.8, 126.6, 125.4, 68.0, 45.2, 14.2.

(S)-2-Amino-1-[3-chloro-4-fluoro-phenyl]ethanol Hydrochloride: wasprepared according to the general procedure outlined above. LRMS[M+H]+189.9; IR (KBr, cm-1) 1509, 3008, 3359; ¹H NMR (500 MHz, DMSO) δ8.21 (b s, 2H), 7.61 (d, J=7.85 Hz, 1H), 7.42 (m, 2H), 6.29 (s, 1H),4.88 (m, 1H), 3.03 (ddd, J=3.4, 12.8, 9.2 Hz, 2H); ¹³C NMR (DMSO, 75.5MHz) δ 157.5, 155.5, 139.7, 128.1, 126.7, 119.3, 116.7, 109.0, 67.8,45.2.

(S)-2-Amino-1-[3-chloro-4-methoxyphenyl]ethanol Hydrochloride: wasprepared according to the general procedure outlined above. LRMS[M+H]+202; IR (KBr, cm-1) 3354, 3003, 2949, 1288, 1064; ¹H NMR (500 MHz,DMSO) δ 8.18 (brs, 3H), 7.43 (d, J=2.0 Hz, 1H), 7.31 (dd, J=8.5, 2.0 Hz,1H), 7.14 (d, J=5.1 Hz, 1H), 6.11 (s, 1H), 4.81 (m, 1H), 3.84 (s, 3H),2.99 (dd, J=13, 3.5 Hz, 1H), 2.83 (dd, J=12.5, 9 Hz, 1H); ¹³C NMR (DMSO,125 MHz) δ 153.9, 135.0, 127.3, 125.8, 120.8, 112.6, 68.0, 56.1, 45.5;Elemental Analysis Calcd for C₉H₁₂ClNO₂-HCl: C, 45.39; H, 5.50; N, 5.88.Found: C, 45.38; H, 5.43; N, 5.70.

(S)-2-Amino-1-(7-bromo-2,3-dihydrobenzfuran-5-yl)-2-aminoethanolHydrochloride: was prepared according to the general procedure outlinedabove. LRMS [M+H]+258; IR (KBr, cm-1) 3349, 3006, 2928, 1485, 1045, 983;¹H NMR (500 MHz, DMSO) δ 8.13 (brs, 3H), 7.29 (s, 1H), 7.23 (s, 1H),6.08 (d, J=4 Hz, 1H), 4.76 (m, 1H), 4.61 (t, J=9 Hz, 2H), 3.29 (t, J=9Hz, 2H), 2.96 (dd, J=13, 3.5 Hz, 1H), 2.82 (dd, J=139.5 Hz, 1H); ¹³C NMR(DMSO, 125 MHz) δ 156.3, 135.9, 129.1, 128.1, 122.1, 100.9, 71.5, 68.2,45.6, 29.9; Elemental Analysis Calcd for C₁₀H₁₂BrNO₂-HCl: C, 40.77; H,4.44; N, 4.75. Found: C, 40.77; H, 4.63; N, 4.63.

General Procedure for the Preparation of2-Amino-3-(substituted-phenyl)-propanol

(S)-[2-(3-Bromo-phenyl)-1-hydroxymethyl-ethyl]-carbamic acid tert-butylester: To a solution of(S)-3-(3-bromo-phenyl)-2-tert-butoxycarbonylamino-propinic acid (500 mg,1.45 mmol) in THF (30 mL) was added borane-tetrahydrofuran complex (1.0M solution) (4.35 mL, 4.35 mmol). The reaction mixture was stirred atroom temperature for 14 h and quenched with acetic acid (1 mL). Afterremoval of most solvent, the residue was extracted with EtOAc, washedwith brine, dried over Na₂SO₄. After concentration, the crude product(400 mg, 83%) was used for the next step without purification. LCMS(M+H)⁺ m/z 330 (t=1.61 min

(S)-2-Amino-3-(3-bromo-phenyl)-propan-1-ol: To a solution of(S)-[2-(3-bromo-phenyl)-1-hydroxymethyl-ethyl]-carbamic acid tert-butylester (400 mg, 1.21 mmol) in methanol (30 mL) was added 4 M HCl indioxane (2 mL, excess). The reaction mixture was stirred at roomtemperature for 14 h. After concentration in vacuo, the residue was usedfor the next step without purification. LCMS (M+H)⁺ m/z 230 (t=0.78min.)

4-Iodo-2-methoxy-pyridine-3-carbaldehyde (WO 95/29917): A 5-literthree-necked round bottom flask was equipped with an overhead mechanicalstirrer under nitrogen, the flask was charged with THF (1 L) and cooledto −78° C. To this stirred solution was added tert-butyllithium (1.7 Msolution in pentane) (800 mL, 1.36 mol) via canula followed by2-methoxypyridine (132.2 g, 1.21 mol) at −78° C. The mixture was stirredfor 1 h at −78° C. To the mixture was addedN-formyl-N,N′,N′-trimethylethylenediamine (176 mL, 1.37 mol) dropwise at−78° C. The reaction mixture was stirred for ca. 30 min at −78° C.before warming to −23° C. over ca. 30 min. To the mixture at −23° C. wasadded ethylene glycol dimethyl ether (1 L) followed by n-butyllithium(2.5 M solution in hexane) (800 mL, 2.0 mol). The resulting mixture wasstirred for ca. 2 h during which time the reaction mixture turned deepgreen. A 12-L 4-necked round flask was charged with iodine (571 g, 2.25mol) and ethylene glycol dimethyl ether (2 L) and the resultant solutionwas cooled to −78° C. The contents of the 5-L flask were transferred viacanula to the mixture of iodine and ethylene glycol dimethyl ether inthe 12-L flask at −78° C. After the addition was complete, the reactionmixture was stirred for an additional 1 h at −78° C. The cooling bathwas removed and the mixture was allowed to warm to about 0° C. andtreated with 2 L of water and 2 L of 1 N hydrochloric acid. Methylt-butyl ether (2 L) was added and the layers were separated. The aqueouslayer was extracted with 2×1 L of methyl t-butyl ether. The combinedorganic layers were washed with saturated Na₂S₂O₃ (1.2 L), brine (1.2L), dried over Na₂SO₄. After concentration in vacuo, the thick slurrywas diluted with hexane (1 L). The mixture was cooled with an ice/waterbath for ca. 30 min. The precipitate was filtered and dried in vacuum toyield the title compound as a light yellow solid. ¹H NMR (300 MHz,CDCl₃) δ 10.22 (s, 1H), 7.86 (1H, d, J=5.3 Hz), 7.54 (1H, d, J=5.3 Hz),4.06 (3H, s). LCMS (M+H)⁺ m/z 364 (t=2.26 min.).

4,6-Dichloro-pyrimidine-5-carbaldehyde: DMF (7 mL, 0.09 mol) was addedto POCl₃ (21 mL, 0.23 mol) at 0° C. The reaction mixture was stirred atroom temperature for 0.5 h. 4,6-Dihydroxy-pyrimidine-5-carbaldehyde (5g, 0.045 mol) was added in small portions. The reaction mixture washeated to 90° C. for 6 h and cooled to room temperature. A large excessof crushed ice was added to the reaction mixture very slowly underice-bath. The mixture was extracted with CH₂Cl₂. The combined organiclayers were washed with water, brine, and dried over Na₂SO₄. Afterconcentration, the residue was purified by column chromatography (20%EtOAc/hexane) to yield the title compound (4 g, 50%). ¹H NMR (400 MHz,CDCl₃) δ 8.91 (1H, s), 7.87 (1H, s). LRMS (M+H)⁺ m/z 177.

6-(4-Iodo-2-methoxy-pyridin-3-yl)-3,5-dihydro-1H-benzo[1,2-d;4,5-d]diimidazol-2-one:A 250 mg (1.0 mml) of the dianiline, and 281 mg (1.0 mml) of theiodoaldehyde were taken in 5 mL of methanol, and the reaction mixturewas stirred at room temperature for 12 hr, solvent was evaporated todryness, and the residue was chromatographed to furnish the product.LRMS [M+H]+408; ¹H NMR (400 MHz, CD₃OD) □ 8.13 (d, J=5.4 Hz, 2H), 7.73(d, J=5.4 Hz, 2H), 7.42 (s, 2H), 3.96 (s, 3H).

6-(4-Chloro-2-oxo-1,2-dihydro-pyridin-3-yl)-3,5-dihydro-1H-benzo[1,2-d;4,5-d]diimidazol-2-one:A 100 mg (0.245 mml) of the methoxy compound was treated with 3 mL of 4N HCl in dioxane, to which was added 0.5 mL of water, and the mixturewas heated at 80° C. for 4 hrs, solvent was evaported to dryness and theresidue was used in the next reaction. LRMS [M+H]+302.

6-(2-Iodo-6-methoxy-phenyl)-1,7-dihydro-1,2,5,7-tetraaza-s-indacene: Toa solution of 1H-indazole-5,6-diamine (250 mg, 1.69 mmol) in methanol(80 mL) was added 2-hydroxy-4-iodo-pyridine-3-carbaldehyde (445 mg, 1.69mmol). The reaction mixture was stirred at room temperature overnight.Concentration in vacuo, the residue was purified by prep. HPLC to yieldthe titled compound (416 mg, 63%). LCMS (t=0.81 min), [M+H]⁺ 392.

4-Chloro-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacene-6-yl)-1H-pyridin-2-one:A suspension of6-(2-iodo-6-methoxy-phenyl)-1,7-dihydro-1,2,5,7-tetraaza-s-indacene (mg,mmol) in 4N HCl dioxane (15 mL) was heated to 80° C. for 4 h.Concentration in vacuo, the residue was used for the next step withoutpurification. LCMS (t=0.76 min), [M+H]⁺ 286.

4-Iodo-2-methoxy-nicotinic acid: To a solution of4-iodo-2-methoxy-pyridine-3-carbaldehyde (4.96 g, 18.9 mmol) in 22 mLtert.-butanol was added in this order 2-methyl-2-butene (30 mL of a 2 Msolution in THF, 60 mmol), natriumdihydrogen-phosphate (5.7 g, 47.5mmol), water (15 mL) and sodiumchlorite (3.9 g, 43 mmol). The mixturewas stirred at ambient temperature for 1 hour, then poured on diluteaqueous formic acid. The mixture was extracted with ethyl acetate, theorganic layer washed with water and brine and concentrated. The residuewas dissolved in diisopropylether+dichloromethane (1+1) and extracted 3times with half-concentrated aqueous NaOH solution. The combined aqueouslayers were acidified with conc. HCl and extracted with ethyl acetate.The organic layers were washed with water and brine, then dried oversodium sulfate and concentrated to give 3.462 g of the title compound asa white solid (67%). ¹H NMR (500 MHz, CD₃OD) δ 7.83 (d, J=5.4 Hz, 1H),7.43 (d, J=5.4 Hz, 1H), 3.94 (s, 3H);

4-Iodo-2-methoxy-nicotinic acid methyl ester: To a solution of4-iodo-2-methoxy-nicotinic acid (1.7 g, 6.1 mmol) in 50 mL of methanol(50 ml) was added trimethylsilyldiazomethane (15 mL of a 2M solution inhexanes, 30 mmol). The mixture was stirred at ambient temperatureovernight, then concentrated. The crude product was purified by flashcolumn chromatography on silica (eluent: hexanes-ethylacetate-triethylamine 100-10-1, then 70-30-1). 1.685 g colorless solidwere isolated (94%). ¹H NMR (500 MHz, acetone-d₆) δ 7.93 (d, J=5.4 Hz,1H), 7.50 (d, J=5.4 Hz, 1H), 3.92 (s, 3H), 3.90 (s, 3H);

N-(4-Amino-pyridin-3-yl)-4-iodo-2-methoxy-nicotinamide (Scheme II): To asuspension of 3,4-diaminopyridine (1.8 g, 16.5 mmol) in 50 mL THF underan atmosphere of argon was added lithiumhexamethyldisilazane (40 mL of a1M solution in hexanes, 40 mmol) and the mixture was stirred for 30minutes at room temperature. A solution of 4-iodo-2-methoxy-nicotinicacid methyl ester (1.8 g, 6.35 mmol) in THF was added and the mixturestirred at room temperature for 4 hours. The mixture was poured on asaturated aqueous solution of ammonium chloride and extracted with ethylacetate. The combined organic layers were washed with water and brine,then dried over sodium sulfate and concentrated. Flash columnchromatography on silica (eluent: chloroform-methanol 100-0, then 95-5,90-10, 80-20) gave 807.8 mg product as an off-white solid. LCMS[M+H]+371, T=0.72 min [YMC ODS-A C18 S7 3.0×50 mm column; 0-100%gradient over 2 min*; 5 mL/min flow rate].*Gradient begins with 10%methanol/90% water (0.1% TFA) and end with 90% methanol/10% water (0.1%TFA); ¹H NMR (500 MHz, DMSO-d₆) δ 9.89 (s, 1H), 8.27 (s, 1H), 7.94 (m,2H), 7.55 (d, J=5.45 Hz, 1H), 6.69 (d, J=5.5 Hz, 1H), 5.76 (s, 2H), 3.91(s, 3H); A NOE enhancement was observed between the amide-NH and C(2)Hof the pyridine, and between the NH2 group and C(5)H of the pyridine.

2-(4-Iodo-2-methoxy-pyridin-3-yl)-1H-imidazo[4,5-c]pyridine (Scheme II):To a solution of N-(4-amino-pyridin-3-yl)-4-iodo-2-methoxy-nicotinamide(612.6 mg, 1.655 mmol) in 10 mL pyridine at 0 C was added (slowly) 1 mLPOCl₃. The mixture was stirred for 2 hours, during which time thecooling bath warmed to 10 C. The mixture was concentrated in vacuum togive 2.0 g of crude product as a black oil that was used as is. LCMS[M+H]+353, T=0.92 min [YMC ODS-A C18 S7 3.0×50 mm column; 0-100%gradient over 2 min*; 5 mL/min flow rate].*Gradient begin with 10%methanol/90% water (0.1% TFA) and end with 90% methanol/10% water (0.1%TFA);

4-Iodo-2-oxo-1,2-dihydro-pyridine-3-carboxylic acid(3-amino-pyridin-4-yl)-amide (Scheme III): A solution of2-(4-iodo-2-methoxy-pyridin-3-yl)-1H-imidazo[4,5-c]pyridine (2.0 gcrude) in 1 M aqueous HCl was refluxed for 17 hours. The mixture waspoured onto a mixture of saturated aqueous sodium bicarbonate solutionand ethyl acetate. The insoluble product was filtered off, rinsed withwater and dried. 346.7 mg of brown powder were isolated (54% over 2steps). MS [M+H]+357, {M−H]−355; ¹H NMR (500 MHz, DMSO-d₆) δ 12.24(broad s, 1H), 9.82 (s, 1H), 8.03 (s, 1H), 7.94 (d, J=5.5 Hz, 1H), 7.23(d, J=6.9 Hz, 1H), 6.72 (d, J=6.9 Hz, 1H), 6.64 (d, J=5.5 Hz, 1H), 6.06(s, 2H); A NOE enhancement was observed between the amide-NH and C(2)Hof the pyridine, and between the NH2 group and C(5)H of the pyridine.

3-(1H-Imidazo[4,5-c]pyridin-2-yl)-4-iodo-1H-pyridin-2-one (Scheme III):To a solution of 4-iodo-2-oxo-1,2-dihydro-pyridine-3-carboxylic acid(4-amino-pyridin-3-yl)-amide (281 mg, 0.79 mmol) in 9 mL pyridine at 0 Cwas added (slowly) 0.5 mL POCl₃ (5.3 mmol). The mixture was stirred at 0C for 2 hours, then poured on a mixture of ice and saturated aqueoussodium bicarbonate solution. The product precipitates as a colloid. Allattempts to filter or extract the product failed. The mixture wasconcentrated in vacuum, water removed by aceotropic destillation withn-propanol and the crude mixture used as it is. LCMS [M+H]+339, T=0.54min product and [M+H]+357, T=0.29 min unreacted starting material [YMCODS-A C18 S7 3.0×50 mm column; 0-100% gradient over 2 min*; 5 mL/minflow rate].*Gradient begins with 10% methanol/90% water (0.1% TFA) andend with 90% methanol/10% water (0.1% TFA)

4-Amino-6-methyl-2-morpholin-4-yl-5-nitropyrimidine: To pressure vessel,a mixture of commercially available4-amino-2-chloro-6-methyl-5-nitropyrimidine (2.00 g, 10.6 mmol) andmorpholine (1.85 mL, 21.2 mmol) in absolute ethanol (40 mL) was stirredat 82° C. for 16 hours. Absolute ethanol was evaporated in vacuo and thecrude product was dissolved in a mixture of THF:EtOAc and this thensubjected on top of a silica gel pad. Elution with EtOAc gave, afterevaporation, the title compound as a yellow solid (2.47 g, 97%): IR(KBr, cm⁻¹) 3468, 3307, 1615, 1560, 1247; ¹H NMR (400 MHz, DMSO) δ 7.96(s, 2H), 3.77 (br s, 4H), 3.62-3.60 (m, 4H), 2.53 (s, 3H); LCMS (⁺ESI,M+H⁺) m/z 240; HPLC: 100% (220 nm).

4-(5-Amino-4-methyl-6-nitro-pyrimidin-2-yl)-piperazine-1-carboxylic acidtert-butyl ester:. To pressure vessel, a mixture of commerciallyavailable 4-amino-2-chloro-6-methyl-5-nitropyrimidine (2.00 g, 10.6mmol) and Boc-piperazine (3.95 g, 21.2 mmol) in absolute ethanol (40 mL)was stirred at 83° C. for 16 hours. Absolute ethanol was evaporated invacuo and the crude product was dissolved in a mixture of THF:EtOAc andthis then subjected on top of a silica gel pad. Elution with EtOAc gave,after evaporation, the title compound as a yellow solid (3.46 g, 96%):IR (KBr, cm⁻¹) 3464, 3327, 2860, 1693, 1571, 1247; ¹H NMR (400 MHz,DMSO) δ 7.98 (s, 2H), 3.79 (br s, 4H), 3.38 (br s, 4H), 2.55 (s, 3H),1.43 (s, 9H); LCMS (⁺ESI, M+H⁺) m/z 283; HPLC: 98% (220 nm).

N⁵-(4-Iodo-2-methoxy-pyridin-3-ylmethylene)-6-methyl-2-morpholin-4-yl-pyrimidine-4,5-diamine:To 4-amino-6-methyl-2-morpholin-4-yl-5nitropyrimidine (500 mg, 2.09mmol) and 10% palladium on carbon (100 mg) were added methanol (10 mL)under nitrogen. The reaction mixture was stirred under hydrogenatmosphere (hydrogen balloon) for 22 hours. The solution was filteredthrough a pad of celite and the filtercake was washed with a smallamount of MeOH. The product in MeOH was used for the next step withoutpurification; LCMS (⁺ESI, M+H⁺) m/z 210; HPLC: 97% (220 nm). To thecrude diamino in MeOH (50 mL) was added a solution of4-iodo-2-methoxy-pyridine-3-carbaldehyde (604 mg, 2.30 mmol) in MeOH (10mL) at 0° C. and stirred for 0.5 hour. The resulting mixture was allowedto warm to ambient temperature and stirred for 6 days in an open systemexposed to air. The reaction mixture was cooled to 0° C. to afford acrystalline yellow solid, which was collected by filtration to give thetitle compound (830 mg, 87%): ¹H NMR (400 MHz, DMSO) δ 8.43 (s, 1H),7.85 and 7.62 (d, J=5.3, 1H), 6.23 (br s, 2H), 3.91 (s, 3H), 3.61 (s,8H), 2.26 (s, 3H); LCMS (⁺ESI, M+H⁺) m/z 455.

8-(4-Iodo-2-methoxy-pyridin-3-yl)-6-methyl-2-morpholin-4-yl-9H-purine:To a solution ofN⁵-(4-Iodo-2-methoxy-pyridin-3ylmethylene)-6-methyl-2-morpholin-4-yl-pyrimidine-4,5-diamine(140 mg, 0.31 mmol) in a mixture of MeOH:THF (20 mL:10 mL) was addediodobenzene diacetate (198 mg, 0.62 mmol). After stirring at ambienttemperature for 4 hours, the solvents were evaporated in vacuo and thecrude was purified by preparative HPLC (see method below) to give thetitle compound as an amber solid (50 mg, 36%): IR (KBr, cm⁻¹) 3420,2853, 1618; ¹H NMR (400 MHz, CDCl₃) δ 9.63 (s, 1H), 7.83 and 7.54 (d,J=5.3, 1H), 3.91 (s, 3H), 3.81-3.77 (m, 8H), 2.72 (s, 3H); LCMS (⁺ESI,M+H⁺) m/z 453; HPLC: 97% (220 nm).

4-[8-(4-Iodo-2-methoxy-pyridin-3-yl)-6-methyl-9H-purin-2-yl]-piperazine-1-carboxylicacid tert-butyl ester: To a stirred solution of4-(5-amino-4-methyl-6-nitro-pyrimidin-2-yl)-piperazine-1-carboxylic acidtert-butyl ester (3.18 g, 9.4 mmol) in methanol (200 mL) and 10%palladium on carbon (180 mg) were added methanol (10 mL) under nitrogen.The reaction mixture was stirred under hydrogen atmosphere (hydrogenballoon) for 18 hours. The solution was filtered through a pad of celiteand the filtercake was washed with a small amount of MeOH. This solutionwas cooled to 0° C. and the 4-iodo-2-methoxy-pyridine-3-carbaldehyde(2.47 g, 9.4 mmol) in methanol (50 mL) was added to the reaction mixtureand stirred at 23° C. for 18 hours. Then iodobenzene diacetate (3.03 g,9.4 mmol) was added and the reaction mixture was stirred at 23° C. for 5hours. MeOH was evaporated in vacuo and the crude material was purifiedon silica gel column using EtOAc:Hex (1:2 to 1:1 gradient) to give thetitle compound as a yellow solid. (2.75 g, 53%). HPLC 98%. LCMS (⁺ESI,M+H⁺) m/z 552; IR (KBr, cm⁻¹) 3177, 2976, 1699, 1558, 1223; ¹H NMR (400MHz, DMSO) δ 12.92 (s, 1H), 8.0 (d, J=5.1 Hz, 1H), 7.64 (d, J=5.6 Hz,1H), 3.81 (s, 3H), 3.73 (br s, 4H), 3.43 (br s, 4H), 2.57 (s, 3H), 1.44(s, 9H).

4-[8-(4-Chloro-2-oxo-1,2-dihydro-pyridin-3-yl)-6-methyl-9H-purin-2-yl]-piperazine-1-carboxylicacid t-butyl ester: To a solution of4-[8-(4-iodo-2-methoxy-pyridin-3-yl)-6-methyl-9H-purin-2-yl]-piperazine-1-carboxylicacid tert-butyl ester (1.4 g, 2.54 mmol) in glacial acetic acid (30 mL)was added 11.6 M HCl (30 mL). The reaction mixture was stirred at 75° C.for 6 hours in a sealed tube and concentrated. The residue was dissolvedin CH₂Cl₂ (250 mL) followed by the addition of triethylamine (2.78 mL,20 mmol). The solution was cooled to 0° C. and di t-butyl dicarbonate(573 mg, 2.63 mmol) was added. The reaction mixture was stirred at 23°C. for 2 hours, poured on saturated NaHCO₃ solution and extracted withCH₂Cl₂ (3×200 mL). The combined organic layers, were dried over MgSO₄and concentrated to give the title compound as a yellow solid. (0.691 g,62%). HPLC 93%; LCMS (⁺ESI, M+H⁺) m/z 446; IR (KBr, cm⁻¹) 3431, 2974,1700, 1506, 1229; ¹H NMR (400 MHz, DMSO+D₂O) δ 7.64 (d, J=7.0 Hz, 0.7H),7.32 (d, J=7.0 Hz, 0.3H), 6.89 (d, J=7.0 Hz, 0.3H), 6.59 (d, J=7.0 Hz,0.7H), 3.75 (br s, 4H), 3.45 (br s, 4H), 2.60 (s, 3H), 1.45 (s, 9H).

6-Chloro-2-methyl-3-nitro-pyridin-4-ylamine (Scheme IX): To a pressurevessel, a degassed solution of 2,6-dichloro-3-nitro-pyridin-4-ylamine(3.0 g, 14.4 mmol) (R. J. Rousseau and R. K. Robins J. HeterocyclicChem. 2, 1965, 196) and tetrakis(triphenylphosphine)palladium(0) (1.67g, 1.44 mmol) in DMF (45 mL) was stirred at room temperature. To thismixture was added dropwise a solution of trimethylaluminium (7.93 mL,15.8 mmol, 2M in toluene). The resulting mixture was sealed and heatedat 70° C. for 3 hours. The reaction mixture was poured into ice/water(800 mL). The organic material was extracted with EtOAc (5×200 mL). Thecombined organic layers were dried (MgSO₄) and concentrated in vacuo.The crude material was purified on silica gel dry column usingEtOAc:Hexane gradient (1:9) to (3:7) to afford the desired compound as ayellow solid (1.20 g, 45%), HPLC: 99% (220 nm), LCMS (⁺ESI, M+H⁺) m/z188, IR (KBr, cm⁻¹) 3431, 3310, 3172, 1653, 1506, 1264; ¹H NMR (400 MHz,CDCl₃) δ 7.19 (s, 1H), 6.57 (br s, 2H), 2.63 (s, 3H);.

2-Methyl-6-morpholin-4-yl-3-nitro-pyridin-4-ylamine (Scheme IX): To apressure vessel, a mixture of6-chloro-2-methyl-3-nitro-pyridin-4-ylamine (0.75 g, 4.00 mmol) andmorpholine (1.40 mL, 16.0 mmol) in absolute ethanol (16 mL) was stirredat 120° C. for 3 days. Absolute ethanol was evaporated in vacuo and thecrude product was dissolved in a mixture of THF:EtOAc and this thensubjected on top of a silica gel pad. Elution with EtOAc gave, afterevaporation, the title compound as a yellow solid (0.89 g, 93%): IR(KBr, cm⁻¹) 3460, 3330, 1627, 1597, 1553, 1233, 1113; ¹H NMR (400 MHz,DMSO) δ 7.13 (s, 2H), 5.89 (s, 1H), 3.66-3.63 (m, 4H), 3.46-3.44 (m,4H), 2.50 (s, 3H); LCMS (⁺ESI, M+H⁺) m/z 239; HPLC: 99% (220 nm).

2-(4-Iodo-2-methoxy-pyridin-3-yl)-4-methyl-6-morpholin-4-yl-1H-imidazo[4,5-c]pyridine(Scheme IX): To 2-methyl-6-morpholin-4-yl-3-nitro-pyridin-4-ylamine (220mg, 0.924 mmol) and 10% palladium on carbon (50 mg) were added methanol(10 mL) under nitrogen. The reaction mixture was stirred under hydrogenatmosphere (hydrogen balloon) for 18 hours. The solution was filteredthrough a pad of celite and the filtercake was washed with a smallamount of MeOH. The product in MeOH was concentrated and used for thenext step without purification; LCMS (⁺ESI, M+H⁺) m/z 209.To the crudediamino in MeOH (5 mL) was added a solution of4-iodo-2-methoxy-pyridine-3-carbaldehyde (267 mg, 1.02 mmol) in MeOH (5mL) at 0° C. and stirred for 0.5 hour. The resulting mixture was allowedto warm to ambient temperature and stirred for 4 days in an open systemexposed to air. The reaction mixture was then heated to 50° C. andstirred for 2 days. MeOH was evaporated in vacuo and the crude waspurified on silica gel column using CH₂Cl₂:MeOH (95:5 gradient) to givethe title compound as a yellow solid (332 mg, 80%): IR (KBr, cm⁻¹) 1624,1554, 1458, 1378; ¹H NMR (400 MHz, DMSO) δ 12.46 (s, 1H), 8.00 and 7.64(d, J=5.5, 1H), 6.54 (s, 1H), 3.80 (s, 3H), 3.74-3.72 (m, 4H), 3.38-3.36(m, 4H), 2.58 (s, 3H); LCMS (⁺ESI, M+H⁺) m/z 452; HPLC: 97% (220 nm).

6-{4-[(S)-2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridin-3-yl}-3,5-dihydro-1H-benzo[1,2-d;4,5-d]diimidazol-2-one:To a stirred solution of 74 mg (0.245 mmol) of the chloropyridone in 2mL of N-methylpyrrolidine was added 3 drops of N-methylmorpholinefollowed by 108 mg (0.612 mmol) of the(S)-2-(3-Chloro-phenyl)-2-hydroxy-ethylamine, and the mixture was heatedat 80° C. for 14 hrs, cooled, and residue was subjected to preparativeHPLC to furnish the product. LRMS [M+H]+437; ¹H NMR (400 MHz, CD₃OD) δ7.27-7.47 (m, 6H), 6.24 (d, J=7.68 Hz. 1H), 4.91 (m, 1H), 3.56 (m, 2H).

4-[(S)-2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-one:The mixture of crude4-chloro-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-onehydrochloric acid salt (80 mg, 0.25 mmol),(S)-2-amino-1-(3-bromo-4-methoxy-phenyl)-ethanol hydrochloric acid salt(105 mg, 0.37 mmol), N-methyl morpholine (0.5 ml, excess), andacetonitrile (15 mL) was heated to 80° C. overnight and cooled to roomtemperature. After concentration in vacuo, the residue was purified byprep. HPLC to yield the titled compound (79 mg, 66%). ¹H NMR (400 MHz,CD₃OD) δ 8.13 (s, 1H), 7.85 (s, 1H), 7.69 (narrow d, J=2.0 Hz, 1H), 7.59(s, 1H), 7.40 (dd, J=2.0, 8.5 Hz, 1H), 7.29 (d, J=7.6 Hz, 1H), 6.96 (d,J=8.5 Hz, 1H), 6.22 (d, J=7.6 Hz, 1H), 4.93 (m, 1H), 3.80 (s, 3H), 3.66(d, J=6.0 Hz, 2H). LCMS (t=1.39 min), [M+H]⁺ 495.

4-[(S)-2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-one:The mixture of crude4-chloro-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-onehydrochloric acid salt (80 mg, 0.25 mmol),(S)-2-amino-1-(3-chloro-phenyl)-ethanol hydrochloric acid salt (80 mg,0.38 mmol), N-methyl morpholine (0.5 ml, excess), and acetonitrile (15mL) was heated to 80° C. overnight and cooled to room temperature. Afterconcentration in vacuo, the residue was purified by prep. HPLC to yieldthe titled compound (64 mg, 61%). ¹H NMR (400 MHz, CD₃OD) δ 8.16 (s,1H), 7.90 (s, 1H), 7.62 (s, 1H), 7.59 (s, 1H), 7.54 (s, 1H), 7.27-7.41(m, 4H), 6.21 (d, J=7.6 Hz, 1H), 4.99 (m, 1H), 3.60-3.71 (m, 2H). LCMS(t=1.44 min), [M+H]⁺ 421.

4-[(S)-2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-one:The mixture of crude4-chloro-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-onehydrochloric acid salt (80 mg, 0.25 mmol),(S)-2-amino-1-(3-bromo-phenyl)-ethanol hydrochloric acid salt (96 mg,0.37 mmol), N-methyl morpholine (0.5 ml, excess), and acetonitrile (15mL) was heated to 80° C. overnight and cooled to room temperature. Afterconcentration in vacuo, the residue was purified by prep. HPLC to yieldthe titled compound (73 mg, 63%). ¹H NMR (400 MHz, CD₃OD) δ 8.08 (s,1H), 7.46 (s, 1H), 7.44 (s, 1H), 7.21-7.40 (m, 5H), 6.14 (d, J=7.5 Hz,1H), 4.97 (m, 1H), 3.60-3.68 (m, 2H). LCMS (t=1.46 min), [M+H]⁺ 465.

4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-onetrifluoroacetate salt: To a suspension of 0.2 mmol of the crude3-(1H-imidazo[4,5-c]pyridin-2-yl)-4-iodo-1H-pyridin-2-one (<0.26 mmol)and 4-iodo-2-oxo-1,2-dihydro-pyridine-3-carboxylic acid(3-amino-pyridin-4-yl)-amide in 3 mL DMF was added 0.1 mL triethylamine(0.72 mmol) and 200 mg 2-(3-chloro-phenyl)-2-triethylsilanyloxy-ethylamine (0.70 mmol). The mixture was stirred under argon at 80 Covernight. Volatile components were evaporated under a stream ofnitrogen. The crude product was purified by preparative HPLC [XTERRAC-18 5 μm, 30×100 mm, 20% solvent B-100% solvent B gradient over 12 min;40 mL/min flow rate, solvent A=10% methanol/90% water (0.1% TFA),solvent B=90% methanol/10% water (0.1% TFA)] to give 3.2 mg pale brownsolid4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-oneLCMS [M+H]+382, T=1.20 min and 9.6 mg brown film4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridine-3-carboxylic acid(4-amino-pyridin-3-yl)-amide LCMS [M+H]+400, T=1.10 min. [YMC ODS-A C18S7 3.0×50 mm column; 0-100% gradient over 2 min*; 5 mL/min flowrate].*Gradient begins with 10% methanol/90% water (0.1% TFA) and endwith 90% methanol/10% water (0.1% TFA);

4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-onetrifluoroacetate: Following the procedure described above using 200 mg2-amino-1-(3-bromo-phenyl)-ethanol (0.93 mmol) gave 15.0 mg4-[2-(3-bromo-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-oneLCMS [M+H]+426, 428, T=1.22 min and 51.7 mg4-[2-(3-bromo-phenyl)-2-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridine-3-carboxylic acid(4-amino-pyridin-3-yl)-amide LCMS [M+H]+444, 446, T=1.14 min. [YMC ODS-AC18 S7 3.0×50 mm column; 0-100% gradient over 2 min*; 5 mL/min flowrate].*Gradient begins with 10% methanol/90% water (0.1% TFA) and endwith 90% methanol/10% water (0.1% TFA); ¹H NMR of imidazo[4,5-c]pyridinproduct (500 MHz, DMSO-d₆) 4:3 mixture of 2 tautomeres “a” and “b” δ14.9 (broad s, 1H a+b), 13.97 (s, 1H, a), 13.86 (s, 1H, b), 11.46 (d,J=6.5 Hz, 1H, b), 11.42 (d, J=6.0 Hz, 1H, a), 10.68 (t, J=5.0 Hz, 1H,b), 10.59 (t, J=5.0 Hz, 1H, a), 9.16 (s, 1H, a), 9.07 (s, 1H, b), 8.52(d, J=6.4 Hz, 1H, a+b), 8.15 (d, J=6.4 Hz, 1H, a), 7.93 (d, J=6.5 Hz,1H, b), 7.72 (s, 1H, a+b), 7.53 (d, J=7.6 Hz, 1H, a+b), 7.46 (m, 2H,a+b) 7.33 (t, J=7.8 Hz, 1H, a+b), 6.26 (d, J=7.6 Hz, 1H, a+b), 6.00(broad s, 1H, a+b), 4.95 (broad s, 1H, a+b), 3.75 (m, 1H, a+b), 3.59 (m,1H, a+b); ¹H NMR of “ring-open amide” product (500 MHz, DMSO-d₆) δ 13.43(broad s, 1H), 12.65 (s, 1H), 11.30 (d, J=6.0 Hz, 1H), 10.52 (t, J=5.3Hz, 1H), 8.51 (s, 1H), 8.09 (d, J=6.8 Hz, 1H), 7.7 (broad s, 2H), 7.64(s, 1H), 7.46 (m, 2H), 7.38, (t, J=6.9 Hz, 1H), 7.31 (t, J=7.83 Hz, 1H),6.97 (d, J=6.8 Hz, 1H), 6.14 (d, J=7.6 Hz, 1H), 5.89 (broad s, 1H), 4.80(dd, J=7.8, 3.8 Hz, 1H), 3.55 (m, 1H), 3.37 (m, 1H).

4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-onetrifluoroacetate salt: Following the procedure described above using 200mg 2-(3-bromo-4-methoxy-phenyl)-2-triethylsilanyloxy-ethylamine (0.55mmol) gave 3.0 mg4-[2-(3-bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-one LCMS [M+H]+456, 458, T=1.16 minand 10.7 mg 4-[2-(3-bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridine-3-carboxylic acid(4-amino-pyridin-3-yl)-amide LCMS [M+H]+474, 476, T=1.07 min. [YMC ODS-AC18 S7 3.0×50 mm column; 0-100% gradient over 2 min*; 5 mL/min flowrate].*Gradient begins with 10% methanol/90% water (0.1% TFA) and endwith 90% methanol/10% water (0.1% TFA).

4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-(6-methyl-2-morpholin-4-yl-9H-purin-8-yl)-1H-pyridin-2-onehydrochloride: To a solution of8-(4-iodo-2-methoxy-pyridin-3-yl)-6-methyl-2-morpholin-4-yl-9H-purine(115 mg, 0.254 mmol) in glacial acetic acid (5 mL) was added 5 mL of11.6 M HCl in a pressure bottle. The vessel was sealed and heated to 85°C. for 15 hours. The reaction mixture was cooled and the vessel openedand concentrated in vacuo to give 117 mg of crude material which wasused for the next step without purification. LCMS (⁺ESI, M+H⁺) m/z 347.To a solution of the crude pyridone (0.254 mmol) in a mixture ofCH₃CN:DMSO (5 mL:1 mL) was added triethylamine (0.35 mL, 2.54 mmol) and(S)-2-amino-1-(3-chloro-phenyl)-ethanol hydrochloride (69 mg, 0.331mmol). The reaction tube was sealed and the mixture stirred at 80° C.for 20 hours. The cooled mixture was concentrated in vacuo and the crudewas purified by preparative HPLC (see method below) to give pureproduct. This solid was dissolved in dioxane and a solution of 1N HCl inEtOH was added to form the hydrochloride salt, which was concentrated.i-PrOH was added to the solid, the solution was cooled to 0° C.,filtrated and dried in vacuo to afford the above title salt as an ambersolid (14 mg, 11%): IR (KBr, cm⁻¹) 3404, 2957, 2855, 1636, 1608, 1494,1231; ¹H NMR (400 MHz, DMSO) δ 11.31 and 10.70 (br s, 1H), 7.56 (s, 1H),7.45 (d, J=7.6, 1H), 7.37-7.29 (m, 4H), 6.20 (d, J=7.5, 1H), 4.92-4.90(m, 1H), 3.69 (s, 8H), 3.55-3.52 (m, 2H), 2.62 (s, 3H); LRMS (⁺ESI,M+H⁺) m/z 482; HPLC: 100% (230 nm); HRMS calcd for C₂₃H₂₄ClN₇O₃:482.1707; found 482.1698.

4-(8-{4-[2-(3-chloro-phenyl)-2(S)-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridin-3-yl}-6-methyl-9H-purin-2-yl)-piperazine-1-carboxylicacid t-butyl ester: To a solution of4-[8-(4-chloro-2-oxo-1,2-dihydro-pyridin-3-yl)-6-methyl-9H-purin-2-yl]-piperazine-1-carboxylicacid t-butyl ester (200 mg, 0.448 mmol) in acetonitrile (10 mL) wasadded triethylamine (125 μl, 0.896 mmol) and(S)-2-amino-1-(3-chloro-phenyl)-ethanol hydrochloride (99 mg, 0.493mmol). The reaction mixture was stirred at 80° C. for 18 hours in asealed tube and concentrated. The residue was dissolved in CH₂Cl₂ (50mL) washed with H₂O (2×30 mL), dried over MgSO₄ and concentrated. Thecrude material was purified by crystallization in hot MeOH (35 mL) togive the title compound as a beige solid. (0.197 g, 76%). HPLC 98%; LCMS(⁺ESI, M+H⁺) m/z 581; IR (KBr, cm⁻¹) 3419, 3271, 2975, 1696, 1496, 1232;¹H NMR (400 MHz, DMSO+D₂O) δ 10.79 (br s, 1H), 7.59-7.32 (m, 5H), 6.22(d, J=7.6 Hz, 1H), 5.00 (br t, 1H), 3.73 (br s, 4H), 3.60 (m, 1H), 3.44(br s, 4H), 2.58 (s, 3H); HRMS calcd for C₂₈H₃₃ClN₈O₄: 581.2391; found581.2389.

4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-(6-methyl-2-piperazin-1-yl-9H-purin-8-yl)-1H-pyridin-2-one:To a solution of4-(8-{4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridin-3-yl}-6-methyl-9H-purin-2-yl)-piperazine-1-carboxylicacid tert-butyl ester (72 mg, 0.124 mmol) in CH₂Cl₂ (30 mL) was addedbenzenesulfonyl fonctionalized silica gel, 0.79 meq/g (7.2 g). Theheterogeneous mixture was stirred at 23° C. for 16 hours. The solventwas removed by filtration and the silica gel was washed with a mixtureof ammonia 2N in MeOH:CH₂Cl₂ (1:1, 20 mL). The ammoniac solution wasevaporated in vacuo to give the title compound as a light yellow solid.(0.053 g, 89%). HPLC 98%; LCMS (⁺ESI, M+H⁺) m/z 481; IR (KBr, cm⁻¹)3262, 2922, 1646, 1597, 1489, 1232; ¹H NMR (400 MHz, DMSO+D₂O) δ 10.80(br s, 1H), 7.59-7.29 (m, 5H), 6.23 (d, J=8.2 Hz, 1H), 5.00 (br t, 1H),3.75 (br s, 4H), 3.62 (m, 1H), 2.89 (br s, 4H), 2.60 (s, 3H); HRMS calcdfor C₂₃H₂₅ClN₈O₂: 480.1789; found 480.1804.

4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-{6-methyl-2-[4-(2-methyl-butyl)-piperazin-1-yl]-9H-purin-8-yl}-1H-pyridin-2-one:To a solution of4-[2-(3-chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-(6-methyl-2-piperazin-1-yl-9H-purin-8-yl)-1H-pyridin-2-one(20 mg, 0.042 mmol) in a mixture of THF:DMF (1.5 mL:0.5 mL) was addedglacial acetic acid (2.3 μl, 0.042 mmol) and 2-methylbutyraldehyde (12μl, 0.117 mmol). The reaction mixture was stirred at 23° C. for 3 hoursfollowed by the addition of NaBH(OAc)₃ (24.9 mg, 0.117 mmol) to thereaction mixture and stirred for an additional 18 hours. MeOH (0.5 mL)was added and the solution was passed through a SCX cartridge (1 g, 0.79meq/g, varian). The cartridge was eluted with MeOH (16 mL) and with 2Nammonia in MeOH (16 mL). The ammonia solution was evaporated in vacuoand the solid obtained was purified using a Shimadzu automatedpreparative HPLC System with the method described below. The solutionwas evaporated on a Savant Speedvac system to give the title compound asa light yellow solid. (14.2 mg, 62%). HPLC 95%; LCMS (⁺ESI, M+H⁺) m/z551; IR (KBr, cm⁻¹) 3403, 2958, 1645, 1616, 1496, 1233; ¹H NMR (400 MHz,DMSO+D₂O) δ 10.78 (brs, 1H), 7.60-7.29 (m, 5H), 6.22 (m, 1H), 4.97 (m,1H), 3.80-3.55 (m, 6H), 2.52-2.15 (m, 8H), 1.75 (brs, 1H), 1.41 (brs,1H), 1.15 (m, 1H) 0.86 (m, 6H).Preparative HPLC Method:

-   Purification Method: Initial gradient (15% B, 85% A) ramp to final    gradient (100% B, 0%A) over 7 minutes, hold for 3 minutes (100% B,    0% A)-   Solvent A: 10% CH₃CN/90% H₂O/5 mmol NH₄OAc-   Solvent B: 10% H₂O/90% CH3₃CN/5 mmol NH₄OAc-   Column: YMC C18 S5 20×100 mm column

The corresponding aldehydes (Examples 12-21) were coupled with4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-(6-methyl-2-piperazin-1-yl-9H-purin-8-yl)-1H-pyridin-2-oneas described previously.

LCMS Exam- Yield Purity (⁺ESI, ple R₁ (%) HPLC M + H⁺) 12

50 >99 575 13

65 >99 561 14

62 >99 572 15

58 >99 578 16 Et 15 >99 509 17

10 >99 615 18

55 >99 535 19

61 >99 607 20

29 >99 574 21

22 >99 589 Example 22 (Scheme IX)

4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-morpholin-4-yl-1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-onebis-hydrochloride salt: To a solution of2-(4-iodo-2-methoxy-pyridin-3-yl)-4-methyl-6-morpholin-4-yl-1H-imidazo[4,5-c]pyridine(75 mg, 0.17 mmol) in glacial acetic acid (3 mL) was added 3 mL of 11.6M HCl in a pressure bottle. The vessel was sealed and heated to 70° C.for 16 hours. The reaction mixture was cooled and the vessel opened andconcentrated in vacuo to give 66 mg of crude material which was used forthe next step without purification. LCMS (⁺ESI, M+H⁺) m/z 346.To asolution of the crude pyridone (0.17 mmol) in a mixture of CH₃CN:DMSO (5mL:1 mL) was added triethylamine (0.23 mL, 1.66 mmol) and(S)-2-amino-1-(3-chloro-phenyl)-ethanol hydrochloride (45 mg, 0.22mmol). The reaction tube was sealed and the mixture stirred at 80° C.for 22 hours. The cooled mixture was concentrated in vacuo and the crudewas purified by preparative HPLC (see method below) to give pure product(51 mg, 64%). A sample (20 mg) was dissolved in dioxane (3 mL) and EtOH(2 mL) and a solution of 1N HCl in EtOH was added to form thebis-hydrochloride salt. The solution was cooled to 0° C., filtrated anddried in vacuo to afford the above title salt as an white solid (19 mg,83%): IR (KBr, cm⁻¹) 3179, 1622, 1532, 1435, 1234; ¹H NMR (400 MHz,DMSO-D₂O) δ 7.53 (br s, 1H), 7.42 (d, J=7.8, 1H), 7.36-7.27 (m, 3H),7.17 (s, 1H), 6.18 (d, J=7.6, 1H), 4.95-4.93 (m, 1H), 3.78 and 3.36 (brt, 8H), 3.74-3.69 (m, 1H), 3.57-3.52 (m, 1H), 2.81 (s, 3H); LRMS (⁺ESI,M+H⁺) m/z 481; HPLC: 100% (230 nm).

1. A compound according to formula I:

its enantiomers, diastereomers, pharmaceutically acceptable salts,hydrates, prodrugs and solvates thereof; wherein A, B, D, and E areeach, independently, C or N provided that if A, B, D, and E are each C,then at least one of R² and R³, R³ and R⁴, or R⁴ and R⁵ is takentogether to form a heterocyclic ring having at least one nitrogen atom;X is selected from the group consisting of N or C wherein each of said Nor C may be optionally substituted, independently, with R⁷ and n is 0,1, 2, or 3; Y is selected from the group consisting of O and S; W isselected from the group consisting of N, C, O, and S, provided that whenW is O or S, R⁴¹ is absent; R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁴¹, and R⁴² areeach independently selected from the group consisting of H, C₁₋₆ alkyl,alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, aminoalkyl,alkoxy, thioalkoxy, nitro, aryl, heteroaryl, alkoxyalkyl,thioalkoxyalkyl, aminoalkyl, aralkyl, heteroarylalkyl,heterocycloalkylalkyl, —CN, —CO₂R⁸, —CONR⁹R¹⁰, —CO₂NR¹¹R¹²,—NR¹³CONR¹⁴R¹⁵, —NR¹⁶SO₂R¹⁷, —SO₂NR¹⁸R¹⁹, —C(NR²⁰)NR²¹R²², —NH-Z,—NH-Z-aryl, and NH-Z-heteroaryl, or any two of R² and R³, R³ and R⁴, orR⁴ and R⁵ can be taken together to form a heterocyclic ring having atleast one nitrogen atom; with the proviso that R³ is not morpholine orsubstituted morpholine; Z is selected from the group consisting of C₁-C₆alkyl, cycloalkyl, alkenyl, cycloalkenyl, and alkynyl; Z optionallyhaving one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo,NR²³SO₂R²⁴ groups and optionally incorporating one or more groupsselected from —CO, —CNOH, —CNOR²⁶, —CNNR²⁷, —CNNCOR²⁸ or —CNNSO₂R²⁹; R⁸,R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹, R²²,R²³, R²⁴, and R²⁶ are independently selected from the group consistingof H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy,alkoxy, aryl, heteroaryl, heterocyclyl, heteroarylalkyl, and alkyl-R²⁵wherein R²⁵ is alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino,alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, heteroaryl,heterocyloalkyl, sulfoxy, sulfonyl, —NR²⁷COOR²⁸, —NR²⁹C(O)R³⁰,—NR³¹SO₂R³², SO₂NR³¹R³², —C(O)NR³³R³⁴, and; R²⁷, R²⁸, R²⁹, R³⁰, R³¹,R³², R³³ and R³⁴ are, independently, hydrogen, alkyl, or cycloalkyl. 2.The compound according to claim 1 wherein R¹, R², R⁴, R⁵, R⁴¹, and R⁴²are absent, H, or C₁ to C₄ alkyl; R³ is H, C₁₋₆ alkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, halo, amino, aminoalkyl, alkoxy,thioalkoxy, nitro, aryl, heteroaryl, alkoxyalkyl, thioalkoxyalkyl,aminoalkyl, aralkyl, heteroarylalkyl, heterocycloalkylalkyl, —CN,—CO₂R⁸, —CONR⁹R¹⁰, —CO₂NR¹¹R¹², —NR¹³CONR¹⁴R¹⁵, —NR¹⁶SO₂R¹⁷,—SO₂NR¹⁸R¹⁹, or —C(NR²⁰)NR²¹R²²; R⁶ is —NH-Z-aryl, or —NH-Z-heteroaryl.3. The compound according to claim 2 wherein R⁶ is —NH-Z-aryl.
 4. Thecompound according to claim 3 wherein said aryl group is a phenyl groupsubstituted at 1 or more positions on the ring.
 5. The compoundaccording to claim 4 wherein said substituent is a Br, F, Cl, or amethoxy group.
 6. The compound according to claim 2 wherein X and E areeach C; W is N; A and D are N; R¹, R⁴¹ and R⁴² and R⁷ are each H; and R⁵is H or CH₃.
 7. The compound according to claim 2 wherein R³ is anoptionally substituted piperazine, homopiperazine, 3-methylpiperazine,3,5-dimethylpiperazine, piperidine, 4-aminopiperidine, imidazole,imidazoline, oxazoline, thiazoline, amide, or amidine.
 8. The compoundaccording to claim 7 wherein R³ is a substituted piperazine.
 9. Thecompound according to claim 2 wherein R⁶ is selected from the groupconsisting of H, 2-aminomethylpyridine, NHCH₂CH(OH)aryl, andNHCH(CH₂OH)CH₂aryl.
 10. The compound according to claim 9 wherein saidaryl is an optionally substituted phenyl.
 11. The compound according toclaim 10 wherein said phenyl is substituted with at least one of Br, Cl,F, alkoxy or —NHSO₂CH₃.
 12. A compound selected from the groupconsisting of:6-{4-[(S)-2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridin-3-yl}-3,5-dihydro-1H-benzo[1,2-d;4,5-d]diimidazol-2-one;4-[(S)-2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-one;4-[(S)-2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-one;4-[(S)-2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-(1,7-dihydro-1,2,5,7-tetraaza-s-indacen-6-yl)-1H-pyridin-2-one;4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-onetrifluoroacetate;4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-onetrifluoroacetate;4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(1H-imidazo[4,5-c]pyridin-2-yl)-1H-pyridin-2-onetrifluoroacetate;4-(8-{4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridin-3-yl}-6-methyl-9H-purin-2-yl)-piperazine-1-carboxylicacid t-butyl ester;4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-(6-methyl-2-piperazin-1-yl-9H-purin-8-yl)-1H-pyridin-2-one;4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-{6-methyl-2-[4-(2-methyl-butyl)-piperazin-1-yl]-9H-purin-8-yl}-1H-pyridin-2-one;4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-methyl-2-[4-(1-methyl-1H-imidazol-2-ylmethyl)-piperazin-1-yl]-9H-purin-8-yl}-1H-pyridin-2-one;4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{2-[4-(1H-imidazol-2-ylmethyl)-piperazin-1-yl]-6-methyl-9H-purin-8-yl}-1H-pyridin-2-one;4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[6-methyl-2-(4-pyridin-2-ylmethyl-piperazin-1-yl)-9H-purin-8-yl]-1H-pyridin-2-one;4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[6-methyl-2-(4-thiazol-2-ylmethyl-piperazin-1-yl)-9H-purin-8-yl]-1H-pyridin-2-one;4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[2-(4-ethyl-piperazin-1-yl)-6-methyl-9H-purin-8-yl]-1H-pyridin-2-one;3-{2-[4-(2-Benzyloxy-ethyl)-piperazin-1-yl]-6-methyl-9H-purin-8-yl}-4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-1H-pyridin-2-one;4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[2-(4-cyclopropylmethyl-piperazin-1-yl)-6-methyl-9H-purin-8-yl]-1H-pyridin-2-one;[4-(8-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-1,2-dihydro-pyridin-3-yl}-6-methyl-9H-purin-2-yl)-piperazin-1-ylmethyl]-cyclopropanecarboxylicacid ethyl ester;4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-methyl-2-[4-(1-methyl-1H-pyrrol-2-ylmethyl)-piperazin-1-yl]-9H-purin-8-yl}-1H-pyridin-2-one;and4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{2-[4-(3-fluoro-benzyl)-piperazin-1-yl]-6-methyl-9H-purin-8-yl}-1H-pyridin-2-one.
 13. Apharmaceutical composition comprising a compound according to claim 1and a pharmaceutically acceptable carrier.
 14. The pharmaceuticalcomposition according to claim 13 further comprising at least one otheranti-cancer agent selected from the group consisting of tamoxifen,toremifen, raloxifene, droloxifene, iodoxyfene, megestrol acetate,anastroxole, letrazole, borazole, exemestane, flutamide, nilutamide,bicalutamide, cyproterone acetate, goserelin acetate, luprolide,finasteride, trastuzumab, methotrexate, 5-fluorouracil, cytosinearabinoside, doxorubicin, daunamycin, epirubicin, idarubicin,mitomycin-C, dacatinomycin, mithramycin, cisplatin, carboplatin,melphalan, chlorambucil, busulphan, cyclophosphamide, ifosfamide,thiotephan, vincristine, paclitaxel, docetaxel, etoposide, teniposide,amsacrine, irinotecan, topotecan, gefitinib, and erlotinib formulated asa fixed dose.
 15. A method of treating a condition associated with atleast one tyrosine kinase enzyme comprising administering to a mammalianspecies in need of such treatment an effective amount of a compoundaccording to claim
 1. 16. The method according to claim 15 wherein saidtyrosine kinase enzyme is Abl, CDK's, EGF, EMT, FGF, FAK, Flk-1/KDR,HER-2, IGF-1R, IR, LCK, MEK, MET, PDGF, Src, or VEGF.
 17. The methodaccording to claim 16 further comprising administering to said mammalianspecies at least one other anti-cancer agent in combination with saidcompound.
 18. The method according to claim 15 wherein the condition iscancer.